Substituted indoles

ABSTRACT

The present invention relates generally to substituted indoles and methods of using them.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.60/505,820 filed Sep. 25, 2003, the entire disclosure of which isincorporated herein by reference.

BACKGROUND

The present invention relates generally to substituted indoles andmethods of using them.

The serine protease inhibitor PAI-1 is one of the primary inhibitors ofthe fibrinolytic system. The fibrinolytic system includes the proenzymeplasminogen, which is converted to the active enzyme, plasmin, by one oftwo tissue type plasminogen activators, t-PA or u-PA. PAI-1 is theprincipal physiological inhibitor of t-PA and u-PA. One of plasmin'smain responsibilities in the fibrinolytic system is to digest fibrin atthe site of vascular injury. The fibrinolytic system, however, is notonly responsible for the removal of fibrin from circulation but is alsoinvolved in several other biological processes including ovulation,embryogenesis, intima proliferation, angiogenesis, tumorigenesis, andatherosclerosis.

Elevated levels of PAI-1 have been associated with a variety of diseasesand conditions including those associated with impairment of thefibrinolytic system. For example, elevated levels of PAI-1 have beenimplicated in thrombotic diseases, e.g., diseases characterized byformation of a thrombus that obstructs vascular blood flow locally ordetaches and embolizes to occlude blood flow downstream. (Krishnamurti,Blood, 69, 798 (1987); Reilly, Arteriosclerosis and Thrombosis, 11, 1276(1991); Carmeliet, Journal of Clinical Investigation, 92, 2756 (1993),Rocha, Fibrinolysis, 8, 294, 1994; Aznar, Haemostasis 24, 243 (1994)).Antibody neutralization of PAI-1 activity resulted in promotion ofendogenous thrombolysis and reperfusion (Biemond, Circulation, 91, 1175(1995); Levi, Circulation 85, 305, (1992)). Elevated levels of PAI-1have also been implicated in diseases such as polycystic ovary syndrome(Nordt, Journal of clinical Endocrinology and Metabolism, 85, 4, 1563(2000)), bone loss induced by estrogen deficiency (Daci, Journal of Boneand Mineral Research, 15, 8, 1510 (2000)), cystic fibrosis, diabetes,chronic periodontitis, lymphomas, diseases associated with extracellularmatrix accumulation, malignancies and diseases associated withneoangiogenesis, inflammatory diseases, vascular damage associated withinfections, and diseases associated with increased uPA levels such asbreast and ovarian cancer.

In view of the foregoing, there exists a need for the identification ofinhibitors of PAI-1 activity and for methods of using the identifiedinhibitors to modulate PAI-1 expression or activity in a subject inorder to treat disorders associated with elevated PAI-1 levels.

SUMMARY

The present invention provides substituted indoles and methods of usingthem. In certain embodiments, substituted 1-aralkyl or 1-aryl-1H-indolesare provided including those of the following formula:

wherein:

-   -   X is a bond or C₁-C₆ alkylene;    -   R₁ and R₂ are, independently, hydrogen, C₁-C₆ alkyl, C₂-C₇        alkenyl, C₂-C₇ alkynyl, C₃-C₈ cycloalkyl, C₇-C₁₁ bicycloalkyl,        C₆-C₁₀ aryl, heterocycle, —C(═O)C₁-C₆ alkyl, —C(═O)aryl,        —C(═O)heterocycle, —C(═O)N(R₆)C₁-C₆ alkyl, —C(═O)N(R₆)aryl,        —C(═O)N(R₆)heterocycle, —SO₂—C₁-C₆ alkyl, —SO₂-aryl, or        —SO₂-heterocycle;    -   or R₁ and R₂ together form a heterocycle;    -   R₃ and R₄ are independently, hydrogen, halogen, C₁-C₆ alkyl,        C₁-C₃ perfluoroalkyl, C₁-C₆ alkoxy, C₃-C₈ cycloalkyl,        —C(═O)C₁-C₃ alkyl, —OH, —NH₂, or —NO₂;    -   R₅ is hydrogen, C₁-C₆ alkyl, C₂-C₇ alkenyl, C₂-C₇ alkynyl, C₃-C₈        cycloalkyl, C₇-C₁₁ bicycloalkyl, C₆-C₁₀ aryl, or heterocyle;    -   A is hydrogen, C₆-C₁₀ aryl, or heterocyle; and    -   R₆ is hydrogen, C₁-C₆ alkyl, halogen, C₂-C₇ alkenyl, C₂-C₇        alkynyl, C₃-C₈ cycloalkyl, aralkyl, C₆-C₁₀ aryl, heterocycle,        hydroxy, C₁-C₆ alkoxy, aryl-oxy, oxo (═O), —CN, —C(═O)H, —CO₂H,        —OCO₂C₁-C₆ alkyl, —CO₂C₁-C₆ alkyl, —CO₂-aryl, —CO₂(C₁-C₆        alkyl)aryl, —OCO₂-aryl, —C(═O)NH₂, —C(═O)NHOH, amino, C₁-C₆        alkylamino, dialkylamino of 1-6 carbons per alkyl moiety,        —NHC(═O)NH—C₁-C₆ alkyl, —NHSO₂—C₁-C₆ alkyl, —NHSO₂-aryl, or        —NHSO₂-heterocycle.

The present invention also provides, inter alia, pharmaceuticallyacceptable salt or ester forms of Formula 1.

The present invention further provides, inter alia, methods of usingsubstituted indoles. In one aspect of the present invention, atherapeutically effective amount of one or more substituted indoles isadministered to a subject in order to treat a PAI-1 related disorder,e.g., by inhibiting PAI-1 activity in the subject. PAI-1 activity isassociated with a number of diseases and conditions. For example, in oneembodiment of the present invention, PAI-1 activity is associated withimpairment of the fibrinolytic system. In other embodiments, PAI-1activity is associated with thrombosis, e.g., venous thrombosis,arterial thrombosis, cerebral thrombosis, and deep vein thrombosis,atrial fibrillation, pulmonary fibrosis, thromboembolic complications ofsurgery, cardiovascular disease, e.g., myocardial ischemia,atherosclerotic plaque formation, chronic obstructive pulmonary disease,renal fibrosis, polycystic ovary syndrome, Alzheimer's disease, orcancer.

DETAILED DESCRIPTION

A. General Overview

The present invention provides compounds that inhibit PAI-1 activity,processes for preparing such compounds, pharmaceutical compositionscontaining such compounds, and methods for using such compounds inmedical therapies. The compounds have properties that are useful for thetreatment, including the prevention and inhibition, of a wide variety ofdiseases and disorders including those involving the production and/oraction of PAI-1. These include disorders resulting from impairment ofthe fibrinolytic system including, but not limited to, thrombosis,coronary heart disease, renal fibrosis, atherosclerotic plaqueformation, pulmonary disease, myocardial ischemia, atrial fibrillation,coagulation syndromes, thromboembolic complications of surgery,peripheral arterial occlusion and pulmonary fibrosis. Other disordersinclude, but are not limited to, polycystic ovary syndrome, Alzheimer'sdisease, and cancer.

The terms “alkyl” and “alkylene,” as used herein, whether used alone oras part of another group, refer to substituted or unsubstitutedaliphatic hydrocarbon chains, the difference being that alkyl groups aremonovalent (i.e., terminal) in nature whereas alkylene groups aredivalent and typically serve as linkers. Both include, but are notlimited to, straight and branched chains containing from 1 to about 12carbon atoms, preferably 1 to about 6 carbon atoms, unless explicitlyspecified otherwise. For example, methyl, ethyl, propyl, isopropyl,butyl, i-butyl and t-butyl are encompassed by the term “alkyl.”Specifically included within the definition of “alkyl” are thosealiphatic hydrocarbon chains that are optionally substituted. Inrepresentative embodiments of the present invention, optionalsubstituents can include C₁-C₆ alkyl, halogen, C₂-C₇ alkenyl, C₂-C₇alkynyl, C₃-C₈ cycloalkyl, aralkyl, aryl optionally substituted with R₇,heterocycle optionally substituted with R₇, hydroxy, C₁-C₆ alkoxy,aryl-oxy, oxo (═O), —CN, —C(═O)H, —CO₂H, —OCO₂C₁-C₆ alkyl, —CO₂C₁-C₆alkyl, —CO₂-aryl, —CO₂(C₁-C₆ alkyl)aryl, —OCO₂-aryl, —C(═O)NH₂,—C(═O)NHOH, amino, alkylamino, dialkylamino, —NHC(═O)NH-C₁-C₆ alkyl,—NHSO₂—C₁-C₆ alkyl, —NHSO₂-aryl, and —NHSO₂-heterocycle.

R₇ is halogen, C₁-C₆ alkoxy, C₁-C₆ alkyl, C₂-C₇ alkenyl, C₂-C₇ alkynyl,hydroxy, —C(═O)C₁-C₇ alkyl, —SO₂—C₁-C₆ alkyl, —CO₂—C₁-C₆ alkyl, oralkoxycarbonylalkyl. In some embodiments, R₇ includes C₂₋₇ acyl.

The carbon number as used in the definitions herein refers to carbonbackbone and carbon branching, but does not include carbon atoms of thesubstituents, such as alkoxy substitutions and the like.

The term “alkenyl”, as used herein, whether used alone or as part ofanother group, refers to a substituted or unsubstituted aliphatichydrocarbon chain and includes, but is not limited to, straight andbranched chains having 2 to about 10 carbon atoms (unless explicitlyspecified otherwise) and containing at least one double bond.Preferably, the alkenyl moiety has 1 or 2 double bonds. Such alkenylmoieties can exist in the E or Z conformations and the compounds of thisinvention include both conformations. Specifically included within thedefinition of “alkenyl” are those aliphatic hydrocarbon chains that areoptionally substituted. In representative embodiments of the presentinvention, optional substituents can include C₁-C₆ alkyl, halogen, C₂-C₇alkenyl, C₂-C₇ alkynyl, C₃-C₈ cycloalkyl, aralkyl, aryl optionallysubstituted with R₇, heterocycle optionally substituted with R₇,hydroxy, C₁-C₆ alkoxy, aryl-oxy, oxo (═O), —CN, —C(═O)H, —CO₂H,—OCO₂C₁-C₆ alkyl, —CO₂C₁-C₆ alkyl, —CO₂-aryl, —CO₂(C₁-C₆ alkyl)aryl,—OCO₂-aryl, —C(═O)NH₂, —C(═O)NHOH, amino, alkylamino, dialkylamino,—NHC(═O)NH—C₁-C₆ alkyl, —NHSO₂—C₁-C₆ alkyl, —NHSO₂-aryl, and—NHSO₂-heterocycle. Heteroatoms, such as O or S attached to an alkenylshould not be attached to a carbon atom that is bonded to a double bond.

The term “acyl”, employed alone or in combination with other terms, isdefined herein as, unless otherwise stated, either an arylalkyl,heteroarylalkyl, (C₂-C₁₀) straight chain, or (C₄-C₁₁) branched-chainmonovalent hydrocarbon moiety; wherein the carbon atom, covalentlylinked to the defined chemical structure, is oxidized to the carbonyloxidation state. Such hydrocarbon moieties may be mono orpolyunsaturated, and may exist in the E or Z configurations. Thecompounds of this invention are meant to include all possible E and Zconfigurations. Examples of acyl moieties include, but are not limitedto, chemical groups such as acetyl, propionyl, butyryl,3,3-dimethylbutyryl, trifluoroacetyl, pivaloyl, hexanoyl, hexenoyl,decanoyl, benzoyl, nicotinyl, isonicotinyl, and homologs, isomers, andthe like.

The term “alkynyl”, as used herein, whether used alone or as part ofanother group, refers to a substituted or unsubstituted aliphatichydrocarbon chain and includes, but is not limited to, straight andbranched chains having 2 to about 10 carbon atoms (unless explicitlyspecified otherwise) and containing at least one triple bond.Preferably, the alkynyl moiety has about 2 to about 7 carbon atoms. Incertain embodiments, the alkynyl can contain more than one triple bondand, in such cases, the alknyl group must contain at least three carbonatoms. In representative embodiments of the present invention, optionalsubstituents can include C₁-C₆ alkyl, halogen, C₂-C₇ alkenyl, C₂-C₇alkynyl, C₃-C₈ cycloalkyl, aralkyl, aryl optionally substituted with R₇,heterocycle optionally substituted with R₇, hydroxy, C₁-C₆ alkoxy,aryl-oxy, oxo (═O), —CN, —C(═O)H, —CO₂H, —OCO₂C₁-C₆ alkyl, —CO₂C₁-C₆alkyl, —CO₂-aryl, —CO₂(C₁-C₆ alkyl)aryl, —OCO₂-aryl, —C(═O)NH₂,—C(═O)NHOH, amino, alkylamino, dialkylamino, —NHC(═O)NH—C₁-C₆ alkyl,—NHSO₂—C₁-C₆ alkyl, —NHSO₂-aryl, and —NHSO₂-heterocycle. Heteroatoms,such as O or S attached to an alkynyl should not be attached to thecarbon that is bonded to a triple bond.

The term “cycloalkyl” as used herein, whether alone or as part ofanother group, refers to a substituted or unsubstituted alicyclichydrocarbon group having 3 to about 20 carbon atoms (unless explicitlyspecified otherwise), preferably 3 to about 8 carbon atoms. Specificallyincluded within the definition of “cycloalkyl” are those alicyclichydrocarbon groups that are optionally substituted. In representativeembodiments of the present invention, optional substituents can includeC₁-C₆ alkyl, halogen, C₂-C₇ alkenyl, C₂-C₇ alkynyl, C₃-C₈ cycloalkyl,aralkyl, aryl optionally substituted with R₇, heterocycle optionallysubstituted with R₇, hydroxy, C₁-C₆ alkoxy, aryl-oxy, oxo (═O), —CN,—C(═O)H, —CO₂H, —OCO₂C₁-C₆ alkyl, —CO₂C₁-C₆ alkyl, —CO₂-aryl, —CO₂(C₁-C₆alkyl)aryl, —OCO₂-aryl, —C(═O)NH₂, —C(═O)NHOH, amino, alkylamino,dialkylamino, —NHC(═O)NH—C₁-C₆ alkyl, —NHSO₂—C₁-C₆ alkyl, —NHSO₂-aryl,and —NHSO₂-heterocycle.

The term “aryl”, as used herein, whether used alone or as part ofanother group, is defined as a substituted or unsubstituted aromatichydrocarbon ring group having 5 to about 50 carbon atoms (unlessexplicitly specified otherwise) with from about 6 to about 10 atomsbeing preferred. The “aryl” group can have a single ring or multiplecondensed rings. The term “aryl” includes, but is not limited to phenyl,α-naphthyl, O-naphthyl, biphenyl, anthryl, tetrahydronaphthyl,fluorenyl, indanyl, biphenylenyl, and acenaphthenyl. Specificallyincluded within the definition of “aryl” are those aromatic groups thatare optionally substituted. For example, in representative embodimentsof the present invention, the, “aryl” groups are optionally substitutedwith from 1 to 5 substituents selected from the group consisting ofacyloxy, hydroxy, aryl, acyl, alkanoyl, alkyl of 1 to 6 carbon atoms,alkoxy of 1 to 6 carbon atoms, alkenyl of 2 to 6 carbon atoms, alkynylof 2 to 6 carbon atoms, substituted alkyl, substituted alkoxy,substituted alkenyl, substituted alkynyl, amino, amino substituted byone or two alkyl groups of from 1 to 6 carbon atoms, aminoacyl,acylamino, azido, cyano, halo, nitro, thioalkoxy of from 1 to 6 carbonatoms, substituted thioalkoxy of from 1 to 6 carbon atoms, andtrihalomethyl. In an exemplary embodiment of the present invention, theoptional substituents include C₁-C₆ alkyl, halogen, C₂-C₇ alkenyl, C₂-C₇alkynyl, C₃-C₈ cycloalkyl, aralkyl, aryl optionally substituted with R₇,heterocycle optionally substituted with R₇, hydroxy, C₁-C₆ alkoxy, C₁-C₆perfluoroalkoxy, aryl-oxy, oxo (═O), —CN, —C(═O)H, —CO₂H, —OCO₂C₁-C₆alkyl, —CO₂C₁-C₆ alkyl, —CO₂-aryl, —CO₂(C₁-C₆ alkyl)aryl, —OCO₂-aryl,—C(═O)NH₂, —C(═O)NHOH, amino, alkylamino, dialkylamino, —NHC(═O)NH—C₁-C₆alkyl, —NHSO₂—C₁-C₆ alkyl, —NHSO₂-aryl, and —NHSO₂-heterocycle.

The term “alkoxy” as used herein, refers to the group R_(a)—O— whereinR_(a) is an alkyl group as defined above. Specifically included withinthe definition of “alkoxy” are those alkoxy groups that are optionallysubstituted.

The term “aryl-oxy” as used herein, refers to the group R_(b)—O— whereinR_(b) is an aryl group as defined above.

The term “alkanoyl” as used herein, refers to the group —C(═O)-alkylgroup wherein alkyl is defined as above. Exemplary alkanoyl groupsinclude, but are not limited to, acetyl (ethanoyl), n-propanoyl,n-butanoyl, 2-methylpropanoyl, n-pentanoyl, 2-methylbutanoyl,3-methylbutanoyl, 2,2-dimethylpropanoyl, heptanoyl, and decanoyl. Thealkyl moieties of alkanoyl groups can be optionally substituted.

The term “arylalkyl” or “aralkyl” refers to the group —R_(a)—R_(b),where R_(a) is an alkyl group as defined above, substituted by R_(b), anaryl group, as defined above. Preferably the alkyl group has from 1 to 6carbon atoms. Examples of arylalkyl moieties include, but are notlimited to, benzyl, 1-phenylethyl, 2-phenylethyl, 3-phenylpropyl,2-phenylpropyl and the like.

The term “alkoxycarbonylalkyl”, as used herein, refers to the groupR_(c)—O—C(═O)R_(c)—, wherein R_(c) is an alkyl group as defined above.Preferably, the alkoxycarbonylalkyl has from 3 to 13 carbon atoms.

The term “alkylamino”, as used herein, refers to the group R_(c)—NH—,wherein R_(c) is an alkyl group as defined above preferably having from1 to 6 carbon atoms.

The term “dialkylamino” as used herein refers to the group —N(C₁-C₆alkyl)₂.

The term “—SO₂—C₁-C₆ alkyl”, as used herein, refers to the group—S(O₂)—R_(a), wherein R_(a) is an alkyl group of 1 to 6 carbons asdefined above.

The term “bicycloalkyl” refers to an optionally substituted, alkyl grouphaving two bridged rings in its structure and having from about 7 toabout 20 carbon atoms (unless explicitly specified otherwise) (and allcombinations and subcombinations of ranges and specific numbers ofcarbon atoms therein), with from about 7 to about 11 carbon atoms beingpreferred. Exemplary bicycloalkyl-ring structures include, but are notlimited to, norbornyl, bornyl, [2.2.2]-bicyclooctyl, cis-pinanyl,trans-pinanyl, camphanyl, iso-bornyl, and fenchyl. Representativesubstituents include, for example, C₁-C₆ alkyl, halogen, C₂-C₇ alkenyl,C₂-C₇ alkynyl, C₃-C₈ cycloalkyl, aralkyl, aryl optionally substitutedwith R₇, heterocycle optionally substituted with R₇, hydroxy, C₁-C₆alkoxy, aryl-oxy, oxo (═O), —CN, —C(═O)H, —CO₂H, —OCO₂C₁-C₆ alkyl,—CO₂C₁-C₆ alkyl, —CO₂-aryl, —CO₂(C₁-C₆ alkyl)aryl, —OCO₂-aryl,—C(═O)NH₂, —C(═O)NHOH, amino, alkylamino, dialkylamino, —NHC(═O)NH—C₁-C₆alkyl, —NHSO₂—C₁-C₆ alkyl, —NHSO₂-aryl, and —NHSO₂-heterocycle.

The term “heterocycle”, as used herein, whether used alone or as part ofanother group, refers to a stable 3 to about 10-member ring containingcarbons atoms and from 1 to 4 heteroatoms selected from the groupconsisting of nitrogen, oxygen, and sulfur. A heterocycle of thisinvention can be either a monocyclic or bicyclic ring system, and can beeither saturated, unsaturated, or partially saturated. A heterocycle canbe optionally fused to a phenyl ring. Heterocycle groups include, butare not limited to, aziridinyl, azetidinyl, 1,4-dioxanyl,hexahydroazepinyl, piperazinyl, piperidinyl, pyrrolidinyl, morpholinyl,thiomorpholinyl, dihydrobenzimidazolyl, dihydrobenzofuranyl,dihydrobenzothienyl, dihydrobenzoxazolyl, dihydrofuranyl,dihydroimidazolyl, dihydroindolyl, dihydroisooxazolyl,dihydroisothiazolyl, dihydrooxadiazolyl, dihydrooxazolyl,dihydropyrrazinyl, dihydropyrazolyl, dihydropyridinyl,dihydropyrimidinyl, dihydropyrrolyl, dihydroquinolinyl,dihydrotetrazolyl, dihydrothiadiazolyl, dihydrothiazolyl,dihydrothienyl, dihydrotriazolyl, dihydroazetidinyl,dihydro-1,4-dioxanyl, tetrahydrofuranyl, tetrahydrothienyl,tetrahydroquinolinyl, and tetrahydroisoquinolinyl. Preferred heterocyclemoieties include: (a) 6-membered saturated, partially unsaturated, orunsaturated heterocycles containing 1-2 nitrogens, optionally fused to aphenyl ring; (b) 5-membered saturated, partially saturated, orunsaturated heterocycles containing 1-3 nitrogen, oxygen, or sulfuratoms, optionally fused to a phenyl ring; (c) saturated, partiallyunsaturated, or unsaturated bicyclic heterocycles containing 1-4nitrogen, oxygen, or sulfur atoms; (d) carbazole, dibenzofuran, anddibenzothiophene. Specifically included in the definition of“heterocycle” are those heterocycles that are optionally substituted.Representative substituents include, for example, C₁-C₆ alkyl, halogen,C₂-C₇ alkenyl, C₂-C₇ alkynyl, C₃-C₈ cycloalkyl, aralkyl, aryl optionallysubstituted with R₇, heterocycle optionally substituted with R₇,hydroxy, C₁-C₆ alkoxy, aryl-oxy, oxo (═O), —CN, —C(═O)H, —CO₂H,—OCO₂C₁-C₆ alkyl, —CO₂C₁-C₆ alkyl, —CO₂-aryl, —CO₂(C₁-C₆ alkyl)aryl,—OCO₂-aryl, —C(═O)NH₂, —C(═O)NHOH, amino, alkylamino, dialkylamino,—NHC(═O)NH—C₁-C₆ alkyl, —NHSO₂—C₁-C₆ alkyl, —NHSO₂-aryl, and—NHSO₂-heterocycle.

The term “heteroaryl” as used herein is defined as a substituted orunsubstituted aromatic heterocyclic ring system (monocyclic orbicyclic). Heteroaryl groups can have, for example, from about 3 toabout 50 carbon atoms (unless explicitly specified otherwise) with fromabout 4 to about 10 being preferred. In some embodiments, heteroarylgroups are aromatic heterocyclic rings systems having about 4 to about14 ring atoms including carbon atoms and 1, 2, 3, or 4 heteroatomsselected from oxygen, nitrogen or sulfur. Representative heteroarylgroups are furan, thiophene, indole, azaindole, oxazole, thiazole,isoxazole, isothiazole, imidazole, N-methylimidazole, pyridine,pyrimidine, pyrazine, pyrrole, N-methylpyrrole, pyrazole,N-methylpyrazole, 1,3,4-oxadiazole, 1,2,4-triazole,1-methyl-1,2,4-triazole, 1H-tetrazole, 1-methyltetrazole, benzoxazole,benzothiazole, benzofuran, benzisoxazole, benzimidazole,N-methylbenzimidazole, azabenzimidazole, indazole, quinazoline,quinoline, and isoquinoline. Bicyclic aromatic heteroaryl goups includephenyl, pyridine, pyrimidine or pyridizine rings that are (a) fused to a6-membered aromatic (unsaturated) heterocyclic ring having one nitrogenatom; (b) fused to a 5- or 6-membered aromatic (unsaturated)heterocyclic ring having two nitrogen atoms; (c) fused to a 5-memberedaromatic (unsaturated) heterocyclic ring having one nitrogen atomtogether with either one oxygen or one sulfur atom; or (d) fused to a5-membered aromatic (unsaturated) heterocyclic ring having oneheteroatom selected from O, N or S. Specifically included within thedefinition of “heteroaryl” are those aromatic groups that are optionallysubstituted. Accordingly, the heteroaryl groups (e.g., pyridinyl)described herein refer to both unsubstituted or substituted groups. Inrepresentative embodiments of the present invention, the, “heteroaryl”groups are optionally substituted with 1 to 5 substituents selected fromthe group consisting of acyloxy, hydroxy, acyl, alkyl of 1 to 6 carbonatoms, alkoxy of 1 to 6 carbon atoms, alkenyl of 2 to 6 carbon atoms,alkynyl of 2 to 6 carbon atoms, substituted alkyl, substituted alkoxy,substituted alkenyl, substituted alkynyl, amino, amino substituted byone or two alkyl groups of from 1 to 6 carbon atoms, aminoacyl,acylamino, azido, cyano, halo, nitro, thioalkoxy of from 1 to 6 carbonatoms, substituted thioalkoxy of from 1 to 6 carbon atoms, andtrihalomethyl.

The term “perfluoroalkyl”, as used herein, whether used alone or as partof another group, refers to a saturated aliphatic hydrocarbon having 1to 6 carbon atoms and two or more fluorine atoms and includes, but isnot limited to, straight or branched chains, such as —CF₃, —CH₂CF₃,—CF₂CF₃ and —CH(CF₃)₂.

The term “halogen” or “halo” refers to chlorine, bromine, fluorine, andiodine.

The term “treating” or “treatment” refers to any indicia of success inamelioration of an injury, pathology, or condition, including anyobjective or subjective parameter such as abatement; remission;diminishing of symptoms or making the injury, pathology, or conditionmore tolerable to the patient; slowing in the rate of degeneration ordecline; making the final point of degeneration less debilitating; orimproving a subject's physical or mental well-being. The treatment oramelioration of symptoms can be based on objective or subjectiveparameters; including the results of a physical examination,neurological examination, and/or psychiatric evaluation. “Treating” or“treatment of a PAI-1 related disorder” includes preventing the onset ofsymptoms in a subject that may be predisposed to a PAI-1 relateddisorder but does not yet experience or exhibit symptoms of the disorder(prophylactic treatment), inhibiting the symptoms of the disorder(slowing or arresting its development), providing relief from thesymptoms or side-effects of the disorder (including palliativetreatment), and/or relieving the symptoms of the disorder (causingregression). Accordingly, the term “treating” includes theadministration of the compounds or agents of the present invention to asubject to prevent or delay, to alleviate, or to arrest or inhibitdevelopment of the symptoms or conditions associated with PAI-1 relateddisorders, e.g., tumor growth associated with cancer. A skilled medicalpractitioner will know how to use standard methods to determine whethera patient is suffering from a disease associated with enhanced levelsand/or activity of PAI-1, e.g., by examining the patient and determiningwhether the patient is suffering from a disease known to be associatedwith elevated PAI-1 levels or activity or by assaying for PAI-1 levelsin blood plasma or tissue of the individual suspected of suffering froma PAI-1 related disease and comparing PAI-1 levels in the blood plasmaor tissue of the individual suspected of suffering from a PAI-1 relateddisease to PAI-1 levels in the blood plasma or tissue of a healthyindividual. Increased PAI-1 levels are indicative of disease.Accordingly, the present invention provides, inter alia, methods ofadministering a compound of the present invention to a subject anddetermining levels of PAI-1 in the subject. The level of PAI-1 in thesubject can be determined before and/or after administration of thecompound.

In healthy individuals, PAI-1 is found at low levels in the plasma (fromabout 5-26 ng/mL), but it is elevated in many PAI-1 related disorders,including, for example, atherosclerosis (Schneiderman J. et. al, ProcNatl Acad Sci 89: 6998-7002, 1992) deep vein thrombosis (Juhan-Vague I,et. al, Thromb Haemost 57: 67-72, 1987), and non-insulin dependentdiabetes mellitus (Juhan-Vague I, et. al, Thromb Haemost 78: 565-660,1997). PAI-1 stabilizes both arterial and venous thrombi, contributingrespectively to coronary arterial occlusion in post-myocardialinfarction (Hamsten A, et. al. Lancet 2:3-9, 1987), and venousthrombosis following post-operative recovery from orthopedic surgery.(Siemens E T, et. al, J Clin Anesthesia 11: 622-629, 1999). Plasma PAI-1is also elevated, for example, in postmenopausal women, and has beenproposed to contribute to the increased incidence of cardiovasculardisease in this population (Koh K et. al, N Engl J Med 336: 683-690,1997).

The term “PAI-1 related disorder or disease” refers to any disease orcondition that is associated with increased or enhanced expression oractivity of PAI-1 or increased or enhanced expression or activity of agene encoding PAI-1. Examples of such increased activity or expressioncan include one or more of the following: activity of the protein orexpression of the gene encoding the protein is increased above the levelof that in normal subjects; activity of the protein or expression of thegene encoding the protein is in an organ, tissue or cell where it is notnormally detected in normal subjects (i.e. spatial distribution of theprotein or expression of the gene encoding the protein is altered);activity of the protein or expression of the gene encoding the proteinis increased when activity of the protein or expression of the geneencoding the protein is present in an organ, tissue or cell for a longerperiod than in a normal subjects (i.e., duration of activity of theprotein or expression of the gene encoding the protein is increased). Anormal or healthy subject is a subject not suffering from a PAI-1related disorder or disease.

The term “pharmaceutically acceptable excipient” means an excipient thatis useful in preparing a pharmaceutical composition that is generallysafe, non-toxic, and desirable, and includes excipients that areacceptable for veterinary use as well as for human pharmaceutical use.Such excipients can be solid, liquid, semisolid, or, in the case of anaerosol composition, gaseous.

“Pharmaceutically acceptable salts and esters” refers to salts andesters that are pharmaceutically acceptable and have the desiredpharmacological properties. Such salts include, for example, salts thatcan be formed where acidic protons present in the compounds are capableof reacting with inorganic or organic bases. Suitable inorganic saltsinclude, for example, those formed with the alkali metals or alkalineearth metals, e.g. sodium and potassium, magnesium, calcium, andaluminum. Suitable organic salts include, for example, those formed withorganic bases such as the amine bases, e.g. ethanolamine,diethanolamine, triethanolamine, tromethamine, N methylglucamine, andthe like. Pharmaceutically acceptable salts can also include acidaddition salts formed from the reaction of basic moieties, such asamines, in the parent compound with inorganic acids (e.g. hydrochloricand hydrobromic acids) and organic acids (e.g. acetic acid, citric acid,maleic acid, and the alkane- and arene-sulfonic acids such asmethanesulfonic acid and benzenesulfonic acid). Pharmaceuticallyacceptable esters include esters formed from carboxy, sulfonyloxy, andphosphonoxy groups present in the compounds, e.g. C₁₋₆ alkyl esters.When there are two acidic groups present, a pharmaceutically acceptablesalt or ester can be a mono-acid-mono-salt or ester or a di-salt orester; and similarly where there are more than two acidic groupspresent, some or all of such groups can be salified or esterified.Compounds named in this invention can be present in unsalified orunesterified form, or in salified and/or esterified form, and the namingof such compounds is intended to include both the original (unsalifiedand unesterified) compound and its pharmaceutically acceptable salts andesters. Also, certain compounds named in this invention can be presentin more than one stereoisomeric form, and the naming of such compoundsis intended to include all single stereoisomers and all mixtures(whether racemic or otherwise) of such stereoisomers.

“Inhibitors,” “activators,” and “modulators” of expression or ofactivity are used to refer to inhibitory, activating, or modulatingmolecules, respectively, identified using in vitro and in vivo assaysfor expression or activity. Inhibitors of the present invention arecompositions that, inhibit expression of PAI-1 or bind to, partially ortotally block stimulation, decrease, prevent, delay activation,inactivate, desensitize, or down regulate the activity of PAI-1. Samplesor assays comprising PAI-1 can be treated with a composition of thepresent invention and compared to control samples without a compositionof the present invention. Control samples (untreated with compositionsof the present invention) can be assigned a relative activity value of100%. In certain embodiments, inhibition of PAI-1 is achieved when theactivity value relative to the control is about 80% or less, optionally50% or 25, 10%, 5% or 1%.

The terms “pharmaceutically acceptable”, “physiologically tolerable” andgrammatical variations thereof, as they refer to compositions, carriers,diluents and reagents, are used interchangeably and represent that thematerials are capable of administration to or upon a human without theproduction of undesirable physiological effects such as nausea,dizziness, gastric upset and the like which would be to a degree thatwould prohibit administration of the compound.

A “therapeutically effective amount” or “pharmaceutically effectiveamount” means the amount that, when administered to a subject, produceseffects for which it is administered. For example, a “therapeuticallyeffective amount,” when administered to a subject to inhibit PAI-1activity, is sufficient to inhibit PAI-1 activity. A “therapeuticallyeffective amount,” when administered to a subject for treating adisease, is sufficient to effect treatment for that disease.

Except when noted, the terms “subject” or “patient” are usedinterchangeably and refer to mammals such as human patients andnon-human primates, as well as experimental animals such as rabbits,rats, and mice, and other animals. Accordingly, the term “subject” or“patient” as used herein means any mammalian patient or subject to whichthe compounds of the invention can be administered. In an exemplaryembodiment of the present invention, to identify subject patients fortreatment according to the methods of the invention, accepted screeningmethods are employed to determine risk factors associated with atargeted or suspected disease or condition or to determine the status ofan existing disease or condition in a subject. These screening methodsinclude, for example, conventional work-ups to determine risk factorsthat are associated with the targeted or suspected disease or condition.These and other routine methods allow the clinician to select patientsin need of therapy using the methods and formulations of the presentinvention.

When any variable occurs more than one time in any constituent or in anyformula, its definition in each occurrence is independent of itsdefinition at every other occurrence. Combinations of substituentsand/or variables are permissible only if such combinations result instable compounds.

B. Substituted Indoles

The present invention provides substituted indoles. Such compounds arepreferably administered to inhibit PAI-1 expression or activity in asubject and, ultimately, to treat diseases or conditions associated withincreased PAI-1 activity in a subject, e.g., a PAI-1 related disorder.

Substituted indoles include those of the following formula:

wherein:

-   -   X is a bond or C₁-C₆ alkylene R₁ and R₂ are, independently,        hydrogen, C₁-C₆ alkyl, C₂-C₇ alkenyl, C₂-C₇ alkynyl, C₃-C₈        cycloalkyl, C₇-C₁₁ bicycloalkyl, C₆-C₁₀ aryl, heterocyle,        —C(═O)C₁-C₆ alkyl, —C(═O)aryl, —C(═O)heterocycle,        —C(═O)N(R₆)C₁-C₆ alkyl, —C(═O)N(R₆)aryl, —C(═O)N(R₆)heterocycle,        —SO₂—C₁-C₆ alkyl, —SO₂-aryl, or —SO₂-heterocycle;    -   or R₁ and R₂ together form a heterocycle;    -   R₃ and R₄ are independently, hydrogen, halogen, C₁-C₆ alkyl,        C₁-C₃ perfluoroalkyl, C₁-C₆ alkoxy, C₃-C₈ cycloalkyl,        —C(═O)C₁-C₃ alkyl, —OH, —NH₂, or —NO₂;    -   R₅ is hydrogen, C₁-C₆ alkyl, C₂-C₇ alkenyl, C₂-C₇ alkynyl, C₃-C₈        cycloalkyl, C₇-C₁₁ bicycloalkyl, C₆-C₁₀ aryl, or heterocycle;    -   A is hydrogen, C₆-C₁₀ aryl, or heterocycle; and    -   R₆ is hydrogen, C₁-C₆ alkyl, halogen, C₂-C₇ alkenyl, C₂-C₇        alkynyl, C₃-C₈ cycloalkyl, aralkyl, C₆-C₁₀ aryl optionally        substituted with R₇, heterocycle optionally substituted with R₇,        hydroxy, C₁-C₆ alkoxy, aryl-oxy, oxo (═O), —CN, —C(═O)H, —CO₂H,        —OCO₂C₁-C₆ alkyl, —CO₂C₁-C₆ alkyl, —CO₂-aryl, —CO₂(C₁-C₆        alkyl)aryl, —OCO₂-aryl, —C(═O)NH₂, —C(═O)NHOH, amino, C₁-C₆        alkylamino, dialkylamino of 1-6 carbons per alkyl moiety,        —NHC(═O)NH—C₁-C₆ alkyl, —NHSO₂—C₁-C₆ alkyl, —NHSO₂-aryl, or        —NHSO₂-heterocycle.

Compounds of the present invention also include prodrugs, stereoisomers,or pharmaceutically acceptable salts or ester forms of Formula 1.

For use in the present invention, R₁ and R₂ can be hydrogen, C₁-C₆alkyl, C₂-C₇ alkenyl, C₂-C₇ alkynyl, C₃-C₈ cycloalkyl, C₇-C₁₁bicycloalkyl, C₆-C₁₀ aryl, heterocycle, —C(═O)C₁-C₆ alkyl, —C(═O)aryl,—C(═O)heterocycle, —C(═O)N(R₆)C₁-C₆ alkyl, —C(═O)N(R₆)aryl,—C(═O)N(R₆)heterocycle, —SO₂—C₁-C₆ alkyl, —SO₂-aryl, or—SO₂-heterocycle. In certain exemplary embodiments of the presentinvention, R₁ or R₂ are, independently, hydrogen, SO₂-alkyl or SO₂-aryloptionally substituted with —OCF₃, aryl, or alkyl. In certain preferredembodiments, R₁ is hydrogen and R₂ is SO₂-methyl or SO₂-phenyloptionally substituted with —OCF₃, phenyl, or tert-butyl. In suchembodiments, R₃, R₄, R₅, R₆, X, and A are as defined herein.

In some exemplary embodiments of the present invention, R₁ and R₂together form a heterocycle. For example, in certain embodiments, R₁ andR₂ together form pyrrole. In such embodiments, R₃, R₄, R₅, R₆, X, and Aare as defined herein.

R₃ and R₄ can be hydrogen, halogen, C₁-C₆ alkyl, C₁-C₃ perfluoroalkyl,C₁-C₆ alkoxy, C₃-C₈ cycloalkyl, —C(═O)C₁-C₃ alkyl, —OH, —NH₂, or —NO₂.In some embodiments of the present invention, R₃ and R₄ are,independently, hydrogen, or alkyl optionally substituted with halogen,—CN, or alkoxy. In certain preferred embodiments, R₃ and R₄ arehydrogen. In such embodiments, R₁, R₂, R₅, R₆, X, and A are as definedherein.

R₅ can be hydrogen, C₁-C₆ alkyl, C₂-C₇ alkenyl, C₂-C₇ alkynyl, C₃-C₈cycloalkyl, C₇-C₁₁ bicycloalkyl, C₆-C₁₀ aryl, or heterocycle. In certainexemplary embodiments of the present invention, R₅ is hydrogen. In suchembodiments, R₁, R₂, R₃, R₄, Rr, X, and A are as defined herein.

R₆ can be C₁-C₆ alkyl, halogen, C₂-C₇ alkenyl, C₂-C₇ alkynyl, C₃-C₈cycloalkyl, aralkyl, C₆-C₁₀ aryl optionally substituted with R₇,heterocycle optionally substituted with R₇, hydroxy, C₁-C₆ alkoxy,aryl-oxy, oxo (═O), —CN, —C(═O)H, —CO₂H, —OCO₂C₁-C₆ alkyl, —CO₂C₁-C₆alkyl, —CO₂-aryl, —CO₂(C₁-C₆ alkyl)aryl, —OCO₂-aryl, —C(═O)NH₂,—C(═O)NHOH, amino, C₁-C₆ alkylamino, dialkylamino of 1-6 carbons peralkyl moiety, —NHC(═O)NH—C₁-C₆ alkyl, —NHSO₂—C₁-C₆ alkyl, —NHSO₂-aryl,or —NHSO₂-heterocycle.

A can be hydrogen, C₆-C₁₀ aryl, or heterocycle. In certain exemplaryembodiments of the present invention, A is hydrogen, phenyl, orthiophene. In such embodiments, R₁, R₂, R₃, R₄, R₅, R₆, and X are asdefined herein.

Preferred compounds of the present invention include those wherein

-   -   X is a bond or unsubstituted C₁-C₆ alkylene;    -   R₁ and R₂ are, independently, hydrogen, C₁-C₆ alkyl, C₂-C₇        alkenyl, C₂-C₇ alkynyl, C₃-C₈ cycloalkyl, C₇-C₁₁ bicycloalkyl,        C₆-C₁₀ aryl, heterocycle, unsubstituted —C(═O)C₁-C₆ alkyl,        unsubstituted —C(═O)aryl, unsubstituted —C(═O)heterocycle,        unsubstituted —C(═O)N(R₆)C₁-C₆ alkyl, unsubstituted        —C(═O)N(R₆)aryl, unsubstituted —C(═O)N(R₆)heterocycle,        unsubstituted —SO₂—C₁-C₆ alkyl, unsubstituted —SO₂-aryl, or        unsubstituted —SO₂-heterocycle;    -   wherein said C₁-C₆ alkyl, C₂-C₇ alkenyl, C₂-C₇ alkynyl, C₃-C₈        cycloalkyl, C₇-C₁₁ bicycloalkyl, C₆-C₁₀ aryl, heterocycle groups        are optionally substituted with unsubstituted C₁-C₆ alkyl,        halogen, unsubstituted C₂-C₇ alkenyl, unsubstituted C₂-C₇        alkynyl, unsubstituted C₃-C₈ cycloalkyl, unsubstituted aralkyl,        hydroxy, unsubstituted C₁-C₆ alkoxy, unsubstituted aryl-oxy, oxo        (═O), —CN, —C(═O)H, —CO₂H, unsubstituted —OCO₂C₁-C₆ alkyl,        unsubstituted CO₂C₁-C₆ alkyl, unsubstituted —CO₂-aryl,        unsubstituted —CO₂(C₁-C₆ alkyl)aryl, unsubstituted —OCO₂-aryl,        —C(═O)NH₂, —C(═O)NHOH, unsubstituted amino, unsubstituted        alkylamino, unsubstituted dialkylamino, unsubstituted        —NHC(═O)NH—C₁-C₆ alkyl, unsubstituted —NHSO₂—C₁-C₆ alkyl,        unsubstituted —NHSO₂-aryl, unsubstituted —NHSO₂-heterocycle,        aryl (optionally substituted with halogen, unsubstituted C₁-C₆        alkoxy, unsubstituted C₁-C₆ alkyl, unsubstituted C₂-C₇ alkenyl,        unsubstituted C₂-C₇ alkynyl, hydroxy, unsubstituted —C(═O)C₁-C₇        alkyl, unsubstituted —SO₂—C₁-C₆ alkyl, unsubstituted —CO₂—C₁-C₆        alkyl, or unsubstituted alkoxycarbonylalkyl) or heterocycle        (optionally substituted with halogen, unsubstituted C₁-C₆        alkoxy, unsubstituted C₁-C₆ alkyl, unsubstituted C₂-C₇ alkenyl,        unsubstituted C₂-C₇ alkynyl, hydroxy, unsubstituted —C(═O)C₁-C₇        alkyl, unsubstituted —SO₂—C₁-C₆ alkyl, unsubstituted —CO₂—C₁-C₆        alkyl, or unsubstituted alkoxycarbonylalkyl);    -   or R₁ and R₂ together form a heterocycle optionally substituted        with unsubstituted C₁-C₆ alkyl, halogen, unsubstituted C₂-C₇        alkenyl, unsubstituted C₂-C₇ alkynyl, unsubstituted C₃-C₈        cycloalkyl, unsubstituted aralkyl, hydroxy, unsubstituted C₁-C₆        alkoxy, unsubstituted aryl-oxy, oxo (═O), —CN, —C(═O)H, —CO₂H,        unsubstituted —OCO₂C₁-C₆ alkyl, unsubstituted CO₂C₁-C₆ alkyl,        unsubstituted —CO₂-aryl, unsubstituted —CO₂(C₁-C₆ alkyl)aryl,        unsubstituted —OCO₂-aryl, —C(═O)NH₂, —C(═O)NHOH, unsubstituted        amino, unsubstituted alkylamino, unsubstituted dialkylamino,        unsubstituted —NHC(═O)NH—C₁-C₆ alkyl, unsubstituted —NHSO₂—C₁-C₆        alkyl, unsubstituted —NHSO₂-aryl, unsubstituted        —NHSO₂-heterocycle, aryl (optionally substituted with halogen,        unsubstituted C₁-C₆ alkoxy, unsubstituted C₁-C₆ alkyl,        unsubstituted C₂-C₇ alkenyl, unsubstituted C₂-C₇ alkynyl,        hydroxy, unsubstituted —C(═O)C₁-C₇ alkyl, unsubstituted        —SO₂—C₁-C₆ alkyl, unsubstituted —CO₂—C₁-C₆ alkyl, or        unsubstituted alkoxycarbonylalkyl) or heterocycle (optionally        substituted with halogen, unsubstituted C₁-C₆ alkoxy,        unsubstituted C₁-C₆ alkyl, unsubstituted C₂-C₇ alkenyl,        unsubstituted C₂-C₇ alkynyl, hydroxy, unsubstituted —C(═O)C₁-C₇        alkyl, unsubstituted —SO₂—C₁-C₆ alkyl, unsubstituted —CO₂—C₁-C₆        alkyl, or unsubstituted alkoxycarbonylalkyl);    -   R₃ and R₄ are independently, hydrogen, halogen, unsubstituted        C₁-C₃ perfluoroalkyl, unsubstituted C₁-C₆ alkoxy, unsubstituted        C₃-C₈ cycloalkyl, unsubstituted —C(═O)C₁-C₃ alkyl, —OH, —NH₂,        —NO₂ or C₁-C₆ alkyl optionally substituted with halogen, —CN, or        C₁-C₃ alkoxy;    -   R₅ is hydrogen, C₁-C₆ alkyl, C₂-C₇ alkenyl, C₂-C₇ alkynyl, C₃-C₈        cycloalkyl, C₇-C₁₁ bicycloalkyl, C₆-C₁₀ aryl, or heterocycle    -   wherein said wherein said C₁-C₆ alkyl, C₂-C₇ alkenyl, C₂-C₇        alkynyl, C₃-C₈ cycloalkyl, C₇-C₁₁ bicycloalkyl, C₆-C₁₀ aryl, or        heterocycle groups are optionally substituted with unsubstituted        C₁-C₆ alkyl, halogen, unsubstituted C₂-C₇ alkenyl, unsubstituted        C₂-C₇ alkynyl, unsubstituted C₃-C₈ cycloalkyl, unsubstituted        aralkyl, hydroxy, unsubstituted C₁-C₆ alkoxy, unsubstituted        aryl-oxy, oxo (═O), —CN, —C(═O)H, —CO₂H, unsubstituted        —OCO₂C₁-C₆ alkyl, unsubstituted CO₂C₁-C₆ alkyl, unsubstituted        —CO₂-aryl, unsubstituted —CO₂(C₁-C₆ alkyl)aryl, unsubstituted        —OCO₂-aryl, —C(═O)NH₂, —C(═O)NHOH, unsubstituted amino,        unsubstituted alkylamino, unsubstituted dialkylamino,        unsubstituted —NHC(═O)NH—C₁-C₆ alkyl, unsubstituted —NHSO₂—C₁-C₆        alkyl, unsubstituted —NHSO₂-aryl, unsubstituted        —NHSO₂-heterocycle, aryl (optionally substituted with halogen,        unsubstituted C₁-C₆ alkoxy, unsubstituted C₁-C₆ alkyl,        unsubstituted C₂-C₇ alkenyl, unsubstituted C₂-C₇ alkynyl,        hydroxy, unsubstituted —C(═O)C₁-C₇ alkyl, unsubstituted        —SO₂—C₁-C₆ alkyl, unsubstituted —CO₂—C₁-C₆ alkyl, or        unsubstituted alkoxycarbonylalkyl) or heterocycle (optionally        substituted with halogen, unsubstituted C₁-C₆ alkoxy,        unsubstituted C₁-C₆ alkyl, unsubstituted C₂-C₇ alkenyl,        unsubstituted C₂-C₇ alkynyl, hydroxy, unsubstituted —C(═O)C₁-C₇        alkyl, unsubstituted —SO₂—C₁-C₆ alkyl, unsubstituted —CO₂—C₁-C₆        alkyl, or unsubstituted alkoxycarbonylalkyl);    -   A is hydrogen, C₆-C₁₀ aryl, or heterocycle    -   wherein said aryl or heterocycle groups are optionally        substituted with unsubstituted C₁-C₆ alkyl, halogen,        unsubstituted C₂-C₇ alkenyl, unsubstituted C₂-C₇ alkynyl,        unsubstituted C₃-C₈ cycloalkyl, unsubstituted aralkyl, hydroxy,        unsubstituted C₁-C₆ alkoxy, unsubstituted aryl-oxy, oxo (═O),        —CN, —C(═O)H, —CO₂H, unsubstituted —OCO₂C₁-C₆ alkyl,        unsubstituted CO₂C₁-C₆ alkyl, unsubstituted —CO₂-aryl,        unsubstituted —CO₂(C₁-C₆ alkyl)aryl, unsubstituted —OCO₂-aryl,        —C(═O)NH₂, —C(═O)NHOH, unsubstituted amino, unsubstituted        alkylamino, unsubstituted dialkylamino, unsubstituted        —NHC(═O)NH—C₁-C₆ alkyl, unsubstituted —NHSO₂—C₁-C₆ alkyl,        unsubstituted —NHSO₂-aryl, unsubstituted —NHSO₂-heterocycle,        aryl (optionally substituted with halogen, unsubstituted C₁-C₆        alkoxy, unsubstituted C₁-C₆ alkyl, unsubstituted C₂-C₇ alkenyl,        unsubstituted C₂-C₇ alkynyl, hydroxy, unsubstituted —C(═O)C₁-C₇        alkyl, unsubstituted —SO₂—C₁-C₆ alkyl, unsubstituted —CO₂—C₁-C₆        alkyl, or unsubstituted alkoxycarbonylalkyl) or heterocycle        (optionally substituted with halogen, unsubstituted C₁-C₆        alkoxy, unsubstituted C₁-C₆ alkyl, unsubstituted C₂-C₇ alkenyl,        unsubstituted C₂-C₇ alkynyl, hydroxy, unsubstituted —C(═O)C₁-C₇        alkyl, unsubstituted —SO₂—C₁-C₆ alkyl, unsubstituted —CO₂—C₁-C₆        alkyl, or unsubstituted alkoxycarbonylalkyl);    -   and R₆ is hydrogen, unsubstituted C₁-C₆ alkyl, halogen,        unsubstituted C₂-C₇ alkenyl, unsubstituted C₂-C₇ alkynyl,        unsubstituted C₃-C₈ cycloalkyl, unsubstituted aralkyl, hydroxy,        unsubstituted C₁-C₆ alkoxy, unsubstituted aryl-oxy, oxo (═O),        —CN, —C(═O)H, —CO₂H, unsubstituted —OCO₂C₁-C₆ alkyl,        unsubstituted CO₂C₁-C₆ alkyl, unsubstituted —CO₂-aryl,        unsubstituted —CO₂(C₁-C₆ alkyl)aryl, unsubstituted —OCO₂-aryl,        —C(═O)NH₂, —C(═O)NHOH, unsubstituted amino, unsubstituted        alkylamino, unsubstituted dialkylamino, unsubstituted        —NHC(═O)NH—C₁-C₆ alkyl, unsubstituted —NHSO₂—C₁-C₆ alkyl,        unsubstituted —NHSO₂-aryl, unsubstituted —NHSO₂-heterocycle,        aryl (optionally substituted with halogen, unsubstituted C₁-C₆        alkoxy, unsubstituted C₁-C₆ alkyl, unsubstituted C₂-C₇ alkenyl,        unsubstituted C₂-C₇ alkynyl, hydroxy, unsubstituted —C(═O)C₁-C₇        alkyl, unsubstituted —SO₂—C₁-C₆ alkyl, unsubstituted —CO₂—C₁-C₆        alkyl, or unsubstituted alkoxycarbonylalkyl) or heterocycle        (optionally substituted with halogen, unsubstituted C₁-C₆        alkoxy, unsubstituted C₁-C₆ alkyl, unsubstituted C₂-C₇ alkenyl,        unsubstituted C₂-C₇ alkynyl, hydroxy, unsubstituted —C(═O)C₁-C₇        alkyl, unsubstituted —SO₂—C₁-C₆ alkyl, unsubstituted —CO₂—C₁-C₆        alkyl, or unsubstituted alkoxycarbonylalkyl).

In certain embodiments of the present invention, substituted indolesinclude the following compounds:

wherein

-   -   R₃, R₄, X, A, and n are as defined above, and    -   Y is —NHSO₂—C₁-C₆ alkyl, —NHSO₂-aryl, —NHSO₂-heterocycle,        pyrrole, —NH-aryl, or —NH-heterocycle.

In representative embodiments of the present invention, the pyrrolegroup is substituted by C₁-C₆ alkyl, halogen, C₂-C₇ alkenyl, C₂-C₇alkynyl, C₃-C₈ cycloalkyl, aralkyl wherein n is from 1 to 6, aryl,heterocycle, hydroxy, C₁-C₆ alkoxy, aryl-oxy, oxo (═O), —CN, —C(═O)H,—CO₂H, —OCO₂C₁-C₆ alkyl, —CO₂C₁-C₆ alkyl, —CO₂-aryl, —CO₂(C₁-C₆alkyl)aryl, —OCO₂-aryl, —C(═O)NH₂, —C(═O)NHOH, amino, alkylamino,dialkylamino, —NHC(═O)NH-C₁-C₆ alkyl, —NHSO₂—C₁-C₆ alkyl, —NHSO₂-aryl,or —NHSO₂-heterocycle.

Exemplary substituted 1-aralkyl or I-aryl-1H-indoles of the presentinvention include, but are not limited to,1-[4-({[4-(trifluoromethoxy)phenyl]sulfonyl}amino)phenyl]-1H-indole-2-carboxylicacid or a pharmaceutically acceptable salt or ester form thereof;1-[4-(1H-pyrrol-1-yl)phenyl]-1H-indole-2-carboxylic acid or apharmaceutically acceptable salt or ester form thereof;1-{4-[(1,1′-biphenyl-4-ylsulfonyl)amino]benzyl}-1H-indole-2-carboxylicacid or a pharmaceutically acceptable salt or ester form thereof;1-(4-{[(4-tert-butylphenyl)sulfonyl]amino}benzyl)-1H-indole-5-carboxylicacid or a pharmaceutically acceptable salt or ester form thereof;1-{4-[(1,1′-biphenyl-4-ylsulfonyl)amino]benzyl}-1H-indole-5-carboxylicacid or a pharmaceutically acceptable salt or ester form thereof;1-(4-{[(4-tert-butylphenyl)sulfonyl]amino}benzyl)-1H-indole-3-carboxylicacid or a pharmaceutically acceptable salt or ester form thereof;1-{4-[(1,1′-biphenyl-4-ylsulfonyl)amino]benzyl}-1H-indole-3-carboxylicacid or a pharmaceutically acceptable salt or ester form thereof;1-(4-{[(4-tert-butylphenyl)sulfonyl]amino}benzyl)-1H-indole-4-carboxylicacid or a pharmaceutically acceptable salt or ester form thereof;1-{4-[(1,1′-biphenyl-4-ylsulfonyl)amino]benzyl}-1H-indole-4-carboxylicacid or a pharmaceutically acceptable salt or ester form thereof;1-{4-[(1,1′-biphenyl-4-ylsulfonyl)amino]benzyl}-1H-indole-6-carboxylicacid or a pharmaceutically acceptable salt or ester form thereof;3-phenyl-1-{4-[(phenylsulfonyl)amino]benzyl})-1H-indole-2-carboxylicacid or a pharmaceutically acceptable salt or ester form thereof;1-{4[(methylsulfonyl)amino]benzyl}-3-phenyl-1H-indole-2-carboxylic acidor a pharmaceutically acceptable salt or ester form thereof;1-{4-[(phenylsulfonyl)amino]benzyl}-3-thien-2-yl-1H-indole-2-carboxylicacid or a pharmaceutically acceptable salt or ester form thereof;1-{4-[(methylsulfonyl)amino]benzyl}-3-thien-2-yl-1H-indole-2-carboxylicacid or a pharmaceutically acceptable salt or ester form thereof;1-[4-(2,5-dimethyl-1H-pyrrol-1-yl)benzyl]-3-phenyl-1H-indole-2-carboxylicacid or a pharmaceutically acceptable salt or ester form thereof;1-(4-anilinobenzyl)-3-phenyl-1H-indole-2-carboxylic acid or apharmaceutically acceptable salt or ester form thereof;1-(4-anilinobenzyl)-3-thien-2-yl-1H-indole-2-carboxylic acid or apharmaceutically acceptable salt or ester form thereof.

The present invention also provides compositions comprising substitutedindoles, including those compounds of formulas 1 and 2 or a stereoisomeror pharmaceutically acceptable salt thereof, and one or morepharmaceutically acceptable carriers, excipients, or diluents. Suchcompositions include pharmaceutical compositions for treating orcontrolling disease states or conditions associated with increased PAI-1activity. In certain embodiments, the compositions comprise mixtures ofone or more substituted indoles.

Certain of the compounds of formulas 1 and 2 contain stereogenic carbonatoms or other chiral elements and thus give rise to stereoisomers,including enantiomers and diastereomers. The present invention includesall of the stereoisomers of formulas 1 and 2, as well as mixtures of thestereoisomers. Throughout this application, the name of the product,where the absolute configuration of an asymmetric center is notindicated, is intended to embrace the individual stereoisomers as wellas mixtures of stereoisomers.

Where an enantiomer is preferred, it can, in some embodiments, beprovided substantially free of the corresponding enantiomer. Thus, anenantiomer substantially free of the corresponding enantiomer refers toa compound that is isolated or separated via separation techniques orprepared free of the corresponding enantiomer. “Substantially free,” asused herein, means that the compound is made up of a significantlygreater proportion of one enantiomer. In preferred embodiments, thecompound is made up of at least about 90% by weight of a preferredenantiomer. In other embodiments of the invention, the compound is madeup of at least about 99% by weight of a preferred enantiomer. Preferredenantiomers can be isolated from racemic mixtures by any method known tothose skilled in the art, including high performance liquidchromatography (HPLC) and the formation and crystallization of chiralsalts, or preferred enantiomers can be prepared by methods describedherein. Methods for the preparation of preferred enantiomers aredescribed, for example, in Jacques, et al., Enantiomers, Racemates andResolutions (Wiley Interscience, New York, 1981); Wilen, S. H., et al.,Tetrahedron 33:2725 (1977); Eliel, E. L. Stereochemistry of CarbonCompounds (McGraw-Hill, NY, 1962); and Wilen, S. H. Tables of ResolvingAgents and Optical Resolutions p. 268 (E. L. Eliel, Ed., Univ. of NotreDame Press, Notre Dame, Ind. 1972).

Exemplary salt forms of the compounds herein include, but are notlimited to, sodium salts and potassium salts. Other exemplary salt formsof these compounds include, but are not limited to, those formed withpharmaceutically acceptable inorganic and organic bases or acids knownin the art. The acids include, for example, acetic, propionic, lactic,citric, tartaric, succinic, fumaric, maleic, malonic, mandelic, malic,phthalic, hydrochloric, hydrobromic, phosphoric, nitric, sulfuric,methanesulfonic, napthalenesulfonic, benzenesulfonic, toluenesulfonic,camphorsulfonic, and similarly known acceptable aids when a compound ofthis invention contains a basic moiety. Salt forms prepared usinginorganic bases include hydroxides, carbonates or bicarbonates of thetherapeutically acceptable alkali metals or alkaline earth metals, suchas sodium potassium, magnesium, calcium and the like. Acceptable organicbases include amines, such as benzylzmine, mono-, di- andtrialkylamines, preferably those having alkyl groups of from 1 to 6carbon atoms, more preferably 1 to 3 carbon atoms, such as methylamine,dimethylamine, trimethylamine, ethylamine, diethylamine, triethylamine,mono-, di-, and triethanolamine. Exemplary salts also include alkylenediamines containing up to 6 carbon atoms, such as hexamethylenediamine;cyclic saturated or unsaturated bases containing up to 6 carbon atoms,including pyrrolidine, peperidine, morpholine, piperazine and theirN-alkyl and N-hydroxyalkyl derivatives, such as N-methyl-morpholine andN-(2-hyroxyethyl)-piperidine, or pyridine. Quaternary salts can also beformed, such as tetralkyl forms, such as tetramethyl forms,alkyl-alkanol forms, such as methyl-triethanol or trimethyl-monoethanolforms, and cyclic ammonium salt forms, such as N-methylpyridinium,N-methyl-N-(2-hydroxyethyl)-morpholinium, N,N-di-methylmorpholinium,N-methyl-N-(2-hydroxyethyl)-morpholinium, or N,N-dimethyl-piperidiniumsalt forms. These salt forms can be prepared using the acidiccompound(s) of formulas 1 or 2 and procedures known in the art.

Exemplary ester forms of the compounds of this invention include, butare not limited to, straight chain alkyl esters having from 1 to 6carbon atoms or branched chain alkyl groups containing 1 to 6 carbonatoms, including methyl, ethyl, propyl, butyl, 2-methylpropyl and1,1-dimethylethyl esters, cycloalkyl esters, alkylaryl esters, benzylesters, and the like. Other exemplary esters include, but are notlimited to, those of the formula —COOR₁₂ wherein R₁₂ is selected fromthe formula:

wherein R₈, R₉, R₁₀, and R₁₁ are independently selected from hydrogen,alkyl of from 1 to 10 carbon atoms, aryl of 6 to 12 carbon atoms,arylalkyl of from 6 to 12 carbon atoms; heteroaryl or alkylheteroarylwherein the heteroaryl ring is bound by an alkyl chain of from 1 to 6carbon atoms.

Preferred compounds of the present invention inhibit PAI-1 activity.Accordingly, the compounds can be used for the treatment, includingprevention, inhibition, and/or amelioration of PAI-1 related disordersin a subject, including, for example, in the treatment of noninsulindependent diabetes mellitus, in the treatment of cardiovascular disease,and in the treatment of thrombotic events associated with coronaryartery and cerebrovascular disease. Using the methods of the presentinvention, a skilled medical practitioner will know how to administersubstituted indoles, including those represented by formulas 1 and 2, toa subject suffering from any of the diseases associated with increasedPAI-1 activity or expression, e.g., diabetes or cardiovascular disease,in order to effect treatment for that disease.

In one exemplary embodiment, substituted indoles are administered to asubject in order to treat disease processes involving thrombotic andprothrombotic states which include, but are not limited to, formation ofatherosclerotic plaques, venous and arterial thrombosis, myocardialischemia, atrial fibrillation, deep vein thrombosis, coagulationsyndromes, pulmonary thrombosis, cerebral thrombosis, thromboemboliccomplications of surgery (such as joint or hip replacement), andperipheral arterial occlusion.

Any disease or condition that is associated with increased PAI-1activity or expression in a subject can be treated using substitutedindoles of the present invention. Exemplary diseases and conditionsinclude stroke, e.g., stroke associated with or resulting from atrialfibrillation; diseases associated with extracellular matrix accumulationincluding, but not limited to, renal fibrosis, chronic obstructivepulmonary disease, polycystic ovary syndrome, restenosis, renovasculardisease, and organ transplant rejection; diseases associated withneoangiogenesis, including, but not limited to, diabetic retinopathy;Alzheimer's disease, e.g., by increasing or normalizing levels ofplasmin concentration in a subject; myelofibrosis with myeloidmetaplasia, e.g., by regulating stromal cell hyperplasia and increasesin extracellular matrix proteins; diabetic nephropathy and renaldialysis associated with nephropathy; malignancies or cancers,including, but not limited to, leukemia, breast cancer and ovariancancer; tumors, including, but not limited to, liposarcomas andepithelial tumors; septicemia; obesity; insulin resistance;proliferative diseases, including, but not limited to, psoriasis;conditions associated with abnormal coagulation homeostasis; low gradevascular inflammation; cerebrovascular diseases; hypertension; dementia;osteoporosis; arthritis; respiratory diseases, such as asthma; heartfailure; arrhythmia; angina, including, but not limited to, anginapectoris; atherosclerosis and sequelae; kidney failure; multiplesclerosis; osteoporosis; osteopenia; dementia; peripheral vasculardisease; peripheral arterial disease; acute vascular syndromes;microvascular diseases including, but not limited to, nephropathy,neuropathy, retinopathy and nephrotic syndrome; hypertension; Type I andII diabetes and related diseases; hyperglycemia; hyperinsulinemia;malignant lesions; premalignant lesions; gastrointestinal malignancies;coronary heart disease, including, but not limited to, primary andsecondary prevention of myocardial infarction, stable and unstableangina, primary prevention of coronary events, and secondary preventionof cardiovascular events; and inflammatory diseases, including, but notlimited to, septic shock and the vascular damage associated withinfections.

The compounds of the present invention can also be administered to asubject in combination with a second therapeutic agent, including, butnot limited to, prothrombolytic, fibrinolytic, and anticoagulant agents,or in conjunction with other therapies, for example, proteaseinhibitor-containing highly active antiretroviral therapy (HAART) forthe treatment of diseases which originate from fibrinolytic impairmentand hyper-coagulability of HIV-1 infected patients. In certainembodiments, the compounds of the present invention can be administeredin conjunction with and/or following processes or procedures involvingmaintaining blood vessel patency, including, but not limited to,vascular surgery, vascular graft and stent patency, organ, tissue andcell implantation and transplantation. The compounds of the presentinvention can also be used for the treatment of blood and blood productsused in dialysis, blood storage in the fluid phase, especially ex vivoplatelet aggregation. The compounds of the present invention can also beadministered to a subject as a hormone replacement agent or to reduceinflammatory markers or C-reactive protein. The compounds can beadministered to improve coagulation homeostasis, to improve endothelialfunction, or as a topical application for wound healing, e.g., theprevention of scarring. The compounds of the present invention can beadministered to a subject in order to reduce the risk of undergoing amyocardial revascularization procedure. The present compounds can alsobe added to human plasma during the analysis of blood chemistry inhospital settings to determine the fibrinolytic capacity thereof. Incertain embodiments, the compounds of the present invention can be usedas imaging agents for the identification of metastatic cancers.

C. Synthesis of Substituted Indoles

Compounds of the present invention can be prepared by those skilled inthe art of organic synthesis employing conventional methods that utilizereadily available reagents and starting materials. Representativecompounds of the present invention can be prepared using the followingsynthetic schemes. The skilled practitioner will know how to make use ofvariants of these process steps, which in themselves are well known inthe art. In the following reaction schemes, R₁, R₂, R₃, R₄, R₅, R₆, andAr (aryl) are as defined above for Formula 1.

The 1H-indole 1 can be alkylated with substituted nitrobenzyl halides inthe presence of a base, such as K₂CO₃, Cs₂CO₃, KOH or NaH, in an inertsolvent, such as THF, dioxane, pyridine, DMF, NMP, or DMSO, at 40 to100° C. The resulting nitro intermediate 2 can be reduced to the aniline3 upon treatment with Raney® nickel in a mixture of hydrazine andethanol at temperature of 0 to 40° C. Compound 3 is then converted tosulfonamide 4, wherein R₁₃ is alkyl, aryl, or Het, upon treatment with asulfonyl halide, preferably alkyl, aryl, or heterocycle sulfonylchloride, and a base such as N,N-diisopropylethylamine in an anhydroussolvent such as dichloromethane for 0.5 to 24 hours at temperature of 0to 40° C. Condensation of aniline 3 with an excess of 1,4-dicarbonylcompounds, such as acetonylacetone, utilizing a Dean-Stark trap forazeotropic water removal leads to pyrrolylindole 6 (Paal-Knorrsynthesis), wherein R₁₄ and R₁₅ are independently hydrogen, alkyl, aryl,or heterocycle. A variant of the Paal-Knorr synthesis which may beapplicable to the synthesis of 1-substituted pyrroles is the reaction ofaniline 3 with an excess of the cyclic acetal2,5-dimethoxytetrahydrofuran. The aniline 3 can also be converted todiaryl amine 8 with boronic acid in the presence of a base, preferably2,6-lutidine, an additive such as myristic acid, and Cu(OAc)₂ in aninert solvent such as toluene at room temperature. The final acids 5, 7or 9 are obtained via a basic hydrolysis of the corresponding ester 4, 6or 8. In this reaction lithium hydroxide, sodium hydroxide, potassiumhydroxide and the like can be used as a base, and water or a mixture ofwater with methanol, ethanol, dioxane and the like can be used as asolvent. The final products can be purified by recrystallization,trituration, preparative thin layer chromatography, flash columnchromatography on silica gel, or high performance liquid chromatography.Purification of intermediates can be achieved in the same manner. A saltis optionally produced by the addition of an acid or base, such ashydrogen chloride gas or hydrochloric acid.

Aryl indole derivatives 12 are also readily prepared by methodsdescribed in the literature or known to those skilled in the art. Forexample, as shown in Scheme II, the 1H-indole 1 can be arylated withsubstituted 4-fluoronitrobenzene in the presence of a base, such Cs₂CO₃,in an inert solvent, such as DMF, at 0 to 40° C. The resulting nitrointermediate 10 can be reduced to the desired aniline 11 upon treatmentwith Raney® nickel in a mixture of hydrazine and ethanol at temperatureof 0 to 40° C. The aniline 11 can be converted to N-aryl indolederivatives 12, such as sulfonamides, pyrroles, and diaryl amines asdescribed in Scheme 1.

The C₃ aryl or heterocycle substituted indole derivative 16, wherein R₁₆is aryl or heterocycle, can be readily prepared as shown in Scheme Im.Substituted indole 1 is first halogenated on the indole C₃ position withknown halogenation agents such as chlorine, bromine,N-chlorosuccinimide, N-bromosuccinimide and the like in a solvent suchas carbon tetrachloride, chloroform, dimethylformamide (DMF), orN-methylpyrrolidinone (NMP) at temperatures of 0 to 30° C. Halo incompound 13 is chlorine, bromine or iodine. The C₃—R₁₆ group can beintroduced by a Suzuki coupling reaction with boronic acid in thepresence of a base, preferably Na₂CO₃, and Pd(PPh₃)₄ in an inert solventsuch as toluene at 80 to 100° C. The 1H-indole intermediate 14 can thenbe alkylated or arylated to afford the desired intermediate 15 which cabto converted to the final product 16 as described in Scheme 1 and Scheme2.

D. Substituted Indoles as Pharmaceutical Compositions

The present invention provides substituted indoles as pharmaceuticals.In a preferred embodiment, substituted indoles are formulated aspharmaceuticals to treat diseases associated with increased PAI-1activity, e.g., by inhibiting PAI-1 activity in a subject.

In general, substituted indoles can be administered as pharmaceuticalcompositions by any method known in the art for administeringtherapeutic drugs including oral, buccal, topical, systemic (e.g.,transdermal, intranasal, or by suppository), or parenteral (e.g.,intramuscular, subcutaneous, or intravenous injection). Compositions cantake the form of tablets, pills, capsules, semisolids, powders,sustained release formulations, solutions, suspensions, emulsions,syrups, elixirs, aerosols, or any other appropriate compositions; andcomprise at least one compound of this invention in combination with atleast one pharmaceutically acceptable excipient. Suitable excipients arewell known to persons of ordinary skill in the art, and they, and themethods of formulating the compositions, can be found in such standardreferences as Alfonso AR: Remington's Pharmaceutical Sciences, 17th ed.,Mack Publishing Company, Easton Pa., 1985. Suitable liquid carriers,especially for injectable solutions, include water, aqueous salinesolution, aqueous dextrose solution, and glycols. In some embodiments ofthe present invention, substituted indoles suitable for use in thepractice of this invention will be administered either singly or incombination with at least one other compound of this invention.Substituted indoles suitable for use in the practice of the presentinvention can also be administered with at least one other conventionaltherapeutic agent for the disease being treated.

Aqueous suspensions of the invention can contain a substituted indole inadmixture with excipients suitable for the manufacture of aqueoussuspensions. Such excipients can include a suspending agent, such assodium carboxymethylcellulose, methylcellulose,hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gumtragacanth and gum acacia, and dispersing or wetting agents such as anaturally occurring phosphatide (e.g., lecithin), a condensation productof an alkylene oxide with a fatty acid (e.g., polyoxyethylene stearate),a condensation product of ethylene oxide with a long chain aliphaticalcohol (e.g., heptadecaethylene oxycetanol), a condensation product ofethylene oxide with a partial ester derived from a fatty acid and ahexitol (e.g., polyoxyethylene sorbitol mono-oleate), or a condensationproduct of ethylene oxide with a partial ester derived from fatty acidand a hexitol anhydride (e.g., polyoxyethylene sorbitan mono-oleate).The aqueous suspension can also contain one or more preservatives suchas ethyl or n-propyl p-hydroxybenzoate, one or more coloring agents, oneor more flavoring agents, and one or more sweetening agents, such assucrose, aspartame or saccharin. Formulations can be adjusted forosmolarity.

Oil suspensions can be formulated by suspending a substituted indole ina vegetable oil, such as arachis oil, olive oil, sesame oil or coconutoil, or in a mineral oil such as liquid paraffin; or a mixture of these.The oil suspensions can contain a thickening agent, such as beeswax,hard paraffin or cetyl alcohol. Sweetening agents can be added toprovide a palatable oral preparation, such as glycerol, sorbitol orsucrose. These formulations can be preserved by the addition of anantioxidant such as ascorbic acid. As an example of an injectable oilvehicle, see Minto, J. Pharmacol. Exp. Ther. 281:93-102, 1997. Thepharmaceutical formulations of the invention can also be in the form ofoil-in-water emulsions. The oily phase can be a vegetable oil or amineral oil, described above, or a mixture of these. Suitableemulsifying agents include naturally-occurring gums, such as gum acaciaand gum tragacanth, naturally occurring phosphatides, such as soybeanlecithin, esters or partial esters derived from fatty acids and hexitolanhydrides, such as sorbitan mono-oleate, and condensation products ofthese partial esters with ethylene oxide, such as polyoxyethylenesorbitan mono-oleate. The emulsion can also contain sweetening agentsand flavoring agents, as in the formulation of syrups and elixirs. Suchformulations can also contain a demulcent, a preservative, or a coloringagent.

The compound of choice, alone or in combination with other suitablecomponents, can be made into aerosol formulations (i.e., they can be“nebulized”) to be administered via inhalation. Aerosol formulations canbe placed into pressurized acceptable propellants, such asdichlorodifluoromethane, propane, nitrogen, and the like.

Formulations suitable for parenteral administration, such as, forexample, by intraarticular (in the joints), intravenous, intramuscular,intradermal, intraperitoneal, and subcutaneous routes, include aqueousand non-aqueous, isotonic sterile injection solutions, which can containantioxidants, buffers, bacteriostats, and solutes that render theformulation isotonic with the blood of the intended recipient, andaqueous and non-aqueous sterile suspensions that can include suspendingagents, solubilizers, thickening agents, stabilizers, and preservatives.Among the acceptable vehicles and solvents that can be employed arewater and Ringer's solution, an isotonic sodium chloride. In addition,sterile fixed oils can conventionally be employed as a solvent orsuspending medium. For this purpose any bland fixed oil can be employedincluding synthetic mono- or diglycerides. In addition, fatty acids suchas oleic acid can likewise be used in the preparation of injectables.These solutions are sterile and generally free of undesirable matter.Where the compounds are sufficiently soluble they can be dissolveddirectly in normal saline with or without the use of suitable organicsolvents, such as propylene glycol or polyethylene glycol. Dispersionsof the finely divided compounds can be made-up in aqueous starch orsodium carboxymethyl cellulose solution, or in suitable oil, such asarachis oil. These formulations can be sterilized by conventional, wellknown sterilization techniques. The formulations can containpharmaceutically acceptable auxiliary substances as required toapproximate physiological conditions such as pH adjusting and bufferingagents, toxicity adjusting agents, e.g., sodium acetate, sodiumchloride, potassium chloride, calcium chloride, sodium lactate and thelike. The concentration of substituted indoles in these formulations canvary widely, and will be selected primarily based on fluid volumes,viscosities, body weight, and the like, in accordance with theparticular mode of administration selected and the patient's needs. ForIV administration, the formulation can be a sterile injectablepreparation, such as a sterile injectable aqueous or oleaginoussuspension. This suspension can be formulated according to the known artusing those suitable dispersing or wetting agents and suspending agents.The sterile injectable preparation can also be a sterile injectablesolution or suspension in a nontoxic parenterally-acceptable diluent orsolvent, such as a solution of 1,3-butanediol. The formulations ofcommends can be presented in unit-dose or multi-dose sealed containers,such as ampules and vials.

Injection solutions and suspensions can be prepared from sterilepowders, granules, and tablets of the kind previously described.

Substituted indoles suitable for use in the practice of this inventioncan be administered orally. The amount of a compound of the presentinvention in the composition can vary widely depending on the type ofcomposition, size of a unit dosage, kind of excipients, and otherfactors well known to those of ordinary skill in the art. In general,the final composition can comprise from, for example, 0.000001 percentby weight (% w) to 10% w of the substituted substituted indole,preferably 0.00001% w to 1% w, with the remainder being the excipient orexcipients.

Pharmaceutical formulations for oral administration can be formulatedusing pharmaceutically acceptable carriers well known in the art indosages suitable for oral administration. Such carriers enable thepharmaceutical formulations to be formulated in unit dosage forms astablets, pills, powder, dragees, capsules, liquids, lozenges, gels,syrups, slurries, suspensions, etc. suitable for ingestion by thepatient. Formulations suitable for oral administration can consist of(a) liquid solutions, such as an effective amount of the packagednucleic acid suspended in diluents, such as water, saline or PEG 400;(b) capsules, sachets or tablets, each containing a predetermined amountof the active ingredient, as liquids, solids, granules or gelatin; (c)suspensions in an appropriate liquid; and (d) suitable emulsions.

Pharmaceutical preparations for oral use can be obtained throughcombination of the compounds of the present invention with a solidexcipient, optionally grinding a resulting mixture, and processing themixture of granules, after adding suitable additional compounds, ifdesired, to obtain tablets or dragee cores. Suitable solid excipientsare carbohydrate or protein fillers and include, but are not limited tosugars, including lactose, sucrose, mannitol, or sorbitol; starch fromcorn, wheat, rice, potato, or other plants; cellulose such as methylcellulose, hydroxymethyl cellulose, hydroxypropylmethyl-cellulose orsodium carboxymethylcellulose; and gums including arabic and tragacanth;as well as proteins such as gelatin and collagen. If desired,disintegrating or solubilizing agents can be added, such as thecross-linked polyvinyl pyrrolidone, agar, alginic acid, or a saltthereof, such as sodium alginate. Tablet forms can include one or moreof lactose, sucrose, mannitol, sorbitol, calcium phosphates, cornstarch, potato starch, microcrystalline cellulose, gelatin, colloidalsilicon dioxide, talc, magnesium stearate, stearic acid, and otherexcipients, colorants, fillers, binders, diluents, buffering agents,moistening agents, preservatives, flavoring agents, dyes, disintegratingagents, and pharmaceutically compatible carriers. Lozenge forms cancomprise the active ingredient in a flavor, e.g., sucrose, as well aspastilles comprising the active ingredient in an inert base, such asgelatin and glycerin or sucrose and acacia emulsions, gels, and the likecontaining, in addition to the active ingredient, carriers known in theart.

The substituted indoles of the present invention can also beadministered in the form of suppositories for rectal administration ofthe drug. These formulations can be prepared by mixing the drug with asuitable non-irritating excipient which is solid at ordinarytemperatures but liquid at the rectal temperatures and will thereforemelt in the rectum to release the drug. Such materials are cocoa butterand polyethylene glycols.

The compounds of the present invention can also be administered byintranasal, intraocular, intravaginal, and intrarectal routes includingsuppositories, insufflation, powders and aerosol formulations (forexamples of steroid inhalants, see Rohatagi, J. Clin. Pharmacol.35:1187-1193, 1995; Tjwa, Ann. Allergy Asthma Immunol. 75:107-111,1995).

The substituted indoles of the present invention can be deliveredtransdermally, by a topical route, formulated as applicator sticks,solutions, suspensions, emulsions, gels, creams, ointments, pastes,jellies, paints, powders, and aerosols.

Encapsulating materials can also be employed with the compounds of thepresent invention and the term “composition” can include the activeingredient in combination with an encapsulating material as aformulation, with or without other carriers. For example, the compoundsof the present invention can also be delivered as microspheres for slowrelease in the body. In one embodiment, microspheres can be administeredvia intradermal injection of drug-containing microspheres, which slowlyrelease subcutaneously (see Rao, J. Biomater Sci. Polym. Ed. 7:623-645,1995; as biodegradable and injectable gel formulations (see, e.g., Gao,Pharm. Res. 12:857-863, 1995); or, as microspheres for oraladministration (see, e.g., Eyles, J. Pharm. Pharmacol. 49:669-674,1997). Both transdermal and intradermal routes afford constant deliveryfor weeks or months. Cachets can also be used in the delivery of thecompounds of the present invention, e.g., anti-atheroscleroticmedicaments.

In another embodiment, the compounds of the present invention can bedelivered by the use of liposomes which fuse with the cellular membraneor are endocytosed, i.e., by employing ligands attached to the liposome,or attached directly to the oligonucleotide, that bind to surfacemembrane protein receptors of the cell resulting in endocytosis. Byusing liposomes, particularly where the liposome surface carries ligandsspecific for target cells, or are otherwise preferentially directed to aspecific organ, one can focus the delivery of the compound into thetarget cells in vivo. (See, e.g., Al-Muhammed, J. Microencapsul.13:293-306, 1996; Chonn, Curr. Opin. Biotechnol. 6:698-708, 1995; Ostro,Am. J. Hosp. Pharm. 46:1576-1587, 1989).

In other cases, the preferred preparation can be a lyophilized powderwhich may contain, for example, any or all of the following: 1 mM-50 mMhistidine, 0.1%-2% sucrose, 2%-7% mannitol, at a pH range of 4.5 to 5.5,that is combined with buffer prior to use.

A pharmaceutical composition of the invention can optionally contain, inaddition to a substituted substituted indole, at least one othertherapeutic agent useful in the treatment of a disease or conditionassociated with increased PAI-1 activity.

The pharmaceutical compositions are generally formulated as sterile,substantially isotonic and in full compliance with all GoodManufacturing Practice (GMP) regulations of the U.S. Food and DrugAdministration.

E. Determining Dosage Regimens for Substituted Indoles

The present invention provides methods of inhibiting PAI-1 activity in asubject for the treatment of diseases and conditions associated withincreased PAI-1 activity using substituted indoles. In an exemplaryembodiment of the present invention, a skilled practitioner will treat asubject having a disease associated with elevated PAI-1 levels and/oractivity with the compounds of the present invention.

For treatment purposes, the compositions or compounds disclosed hereincan be administered to the subject in a single bolus delivery, viacontinuous delivery (e.g., continuous transdermal, mucosal, orintravenous delivery) over an extended time period, or in a repeatedadministration protocol (e.g., by an hourly, daily or weekly, repeatedadministration protocol). The pharmaceutical formulations of the presentinvention can be administered, for example, one or more times daily, 3times per week, or weekly. In an exemplary embodiment of the presentinvention, the pharmaceutical formulations of the present invention areorally administered once or twice daily.

In this context, a therapeutically effective dosage of the biologicallyactive agent(s) can include repeated doses within a prolonged treatmentregimen that will yield clinically significant results to alleviate oneor more symptoms or detectable conditions associated with increasedPAI-1 activity. Determination of effective dosages in this context istypically based on animal model studies followed up by human clinicaltrials and is guided by determining effective dosages and administrationprotocols that significantly reduce the occurrence or severity oftargeted exposure symptoms or conditions in the subject. Suitable modelsin this regard include, for example, murine, rat, porcine, feline,non-human primate, and other accepted animal model subjects known in theart. Alternatively, effective dosages can be determined using in vitromodels (e.g., immunologic and histopathologic assays). Using suchmodels, only ordinary calculations and adjustments are typicallyrequired to determine an appropriate concentration and dose toadminister a therapeutically effective amount of the biologically activeagent(s) (e.g., amounts that are intranasally effective, transdermallyeffective, intravenously effective, or intramuscularly effective toelicit a desired response). In alternative embodiments, an “effectiveamount” or “therapeutically effective dose” of the biologically activeagent(s) will simply inhibit or enhance one or more selected biologicalactivity(ies) correlated with a disease or condition, as set forthabove, for either therapeutic or diagnostic purposes.

The actual dosage of biologically active agents will of course varyaccording to factors such as the extent of exposure and particularstatus of the subject (e.g., the subject's age, size, fitness, extent ofsymptoms, susceptibility factors, etc), time and route ofadministration, as well as other drugs or treatments being administeredconcurrently. Dosage regimens can be adjusted to provide an optimumprophylactic or therapeutic response. By “therapeutically effectivedose” herein is meant a dose that produces effects for which it isadministered. More specifically, a therapeutically effective dose of thecompound(s) of the invention preferably alleviates symptoms,complications, or biochemical indicia of diseases associated withincreased PAI-1 activity. The exact dose will depend on the purpose ofthe treatment, and will be ascertainable by one skilled in the art usingknown techniques (see, e.g., Lieberman, Pharmaceutical Dosage Forms(Vols. 1-3, 1992); Lloyd, 1999, The Art, Science, and Technology ofPharmaceutical Compounding; and Pickar, 1999, Dosage Calculations). Atherapeutically effective dose is also one in which any toxic ordetrimental side effects of the active agent is outweighed in clinicalterms by therapeutically beneficial effects. It is to be further notedthat for each particular subject, specific dosage regimens should beevaluated and adjusted over time according to the individual need andprofessional judgment of the person administering or supervising theadministration of the compound.

In an exemplary embodiment of the present invention, unit dosage formsof the compounds are prepared for standard administration regimens. Inthis way, the composition can be subdivided readily into smaller dosesat the physicians direction. For example, unit dosages can be made up inpacketed powders, vials or ampoules and preferably in capsule or tabletform. The active compound present in these unit dosage forms of thecomposition can be present in an amount of, for example, from about onegram to about fifteen grams or more, for single or multiple dailyadministration, according to the particular need of the patient. Byinitiating the treatment regimen with a minimal daily dose of about onegram, the blood levels of PAI-1 and the patients symptomatic reliefanalysis can be used to determine whether a larger or smaller dose isindicated. Effective administration of the compounds of this inventioncan be given at an oral dose of from, for example about 0.1 mg/kg/day toabout 1,000 mg/kg/day. Preferably, administration will be from about10/mg/kg/day to about 600 mg/kg/day, more preferably from about 25 toabout 200 mg/kg/day, and even more preferably from about 50 mg/kg/day toabout 100 mg/kg/day.

In certain embodiments, the present invention is directed to prodrugs ofcompounds of formulas 1 and 2. The term “prodrug,” as used herein, meansa compound that is convertible in vivo by metabolic means (e.g. byhydrolysis) to a compound of formula 1 or 2. Various forms of prodrugsare known in the art such as those discussed in, for example, Bundgaard,(ed.), Design of Prodrugs, Elsevier (1985); Widder, et al. (ed.),Methods in Enzymology, vol. 4, Academic Press (1985); Krogsgaard-Larsen,et al., (ed). “Design and Application of Prodrugs, Textbook of DrugDesign and Development, Chapter 5, 113-191 (1991), Bundgaard, et al.,Journal of Drug Delivery Reviews, 8:1-38(1992), Bundgaard, J. ofPharmaceutical Sciences, 77:285 et seq. (1988); and Higuchi and Stella(eds.) Prodrugs as Novel Drug Delivery Systems, American ChemicalSociety (1975).

F. Kits

After a pharmaceutical comprising a substituted substituted indole hasbeen formulated in a suitable carrier, it can be placed in anappropriate container and labeled for treatment of a PAI-1 relateddisorder, e.g., leukemia. Additionally, another pharmaceuticalcomprising at least one other therapeutic agent useful in the treatmentof the PAI-1 related disorder can be placed in the container as well andlabeled for treatment of the indicated disease. Alternatively, a singlepharmaceutical comprising a substituted substituted indole and at leastone other therapeutic agent useful in the treatment of a PAI-1 relateddisorder can be placed in an appropriate container and labeled fortreatment. For administration of pharmaceuticals comprising substitutedindoles and of pharmaceuticals comprising, in a single pharmaceutical,substituted indoles and at least one other therapeutic agent useful inthe treatment of a PAI-1 related disorder, such labeling would include,for example, instructions concerning the amount, frequency and method ofadministration. Similarly, for administration of multiplepharmaceuticals provided in the container, such labeling would include,for example, instructions concerning the amount, frequency and method ofadministration of each pharmaceutical.

EXAMPLES

The syntheses of compounds 1-17 are described in examples 1-17respectively.

Example 1 Synthesis of1-[4-({[4-(trifluoromethoxy)phenyl]sulfonyl}amino)phenyl]-1H-indole-2-carboxylicAcid

Step 1: A large excess of Raney® nickel was added in portions to astirred solution of 1-(4-nitrophenyl)-1H-indole-2-carboxylic acid (1.40g, 5.0 mmol), hydrazine (1.0 mL, 32 mmol) in 20 mL of ethanol. Afterstirring at room temperature for 2 h the catalyst was then removed byfiltering through a short pad of Celite®521. The filtrate wasconcentrated to give 1-(4-aminophenyl)-1H-indole-2-carboxylic acid (1.0g, 80%) as a beige solid: MS (ESI) m/z 253 (MH⁺); MS (ESI) m/z 251([M−H]⁻).

Step 2: To a stirred solution of1-(4-aminophenyl)-1H-indole-2-carboxylic acid (0.4 g, 1.59 mmol) in 10mL of 1:1 of dichloromethane/sat. aqueous sodium bicarbonate was added4-trifluoromethoxy-benzenesulfonyl chloride (0.40 g, 1.53 mmol). Afterbeing stirred at room temperature for 1 hour the solid was collected andpurified by semi-preparative HPLC (Column: Phenomenex C18 Luna 21.6mm×60 mm, 5 μM; Solvent A: Water (0.1% TFA buffer); Solvent B:Acetonitrile (0.1% TFA buffer); Solvent Gradient: Time 0: 0% B; 10 min:100% B; Hold 100% B 5 min. Flow Rate: 22.5 mL/min) to afford 0.105 g(15%) of the title compound as a pale yellow solid: ¹H NMR (DMSO-d₆) δ6.89(d, J=8.3 Hz, 1H), 7.10-7.30 (m, 6H), 7.39 (d, J=2.1 Hz, 1H), 7.60(d, J=7.2 Hz, 2H), 7.71 (d, J=6.0 Hz, 1H), 7.90-8.00 (m, 2H), 10.63 (brs, 1H), 12.74 (br s, 1H); MS (ESI) m/z 477 (MH⁺); MS (ESI) m/z 475([M−H]⁻); HRMS calcd for C₂₂H₁₆F₃N₂O₅S: 477.0721; found (ESI⁺):477.0719; Anal. calcd for C₂₂H₁₅F₃N₂O₅S_(0.2)H₂O: C, 55.05; H, 3.23; N,5.84. Found: C, 54.97; H, 3.41; N, 5.82.

Example 2 Synthesis of1-[4-(1H-pyrrol-1-yl)phenyl]-1H-indole-2-carboxylic Acid

A solution of 1-(4-aminophenyl)-1H-indole-2-carboxylic acid (0.4 g, 1.59mmol) and 2,5-dimethoxytetrahydrofuran (0.50 g, 3.78 mmol) in 15 mL ofglacial acetic acid was heated at 90° C. for 1.5 hours. After removingthe solvent the residue was purified by by semi-preparative HPLC toafford the title compound (0.095 mg, 20%) as a beige solid: ¹H NMR(DMSO-d₆) δ 6.32 (s, 2H), 7.08 (d, J=8.4 Hz, 1H), 7.15-7.25 (m, 1H),7.25-7.35 (m, 1H), 7.40-7.55 (m, 5H), 7.60-7.70 (m, 3H), 12.82 (br s,1H); MS (ESI) m/z 303 (MH⁺); MS (ESI) m/z 301 ([M−H]⁻); HRMS calcd forC₁₉H₁₅N₂O₂: 303.1122; found (ESI⁺): 303.1124; Anal. calcd forC₁₉H₁₄N₂O₂.0.1H₂O: C, 75.0; H, 4.71; N, 9.21. Found: C, 74.90; H, 4.42;N, 9.33.

Example 3 Synthesis of1-{4-[(1,1′-biphenyl-4-ylsulfonyl)amino]benzyl}-1H-indole-2-carboxylicAcid

Step 1: A mixture of 4-nitrobenzyl bromide (1.85 g, 8.56 mmol),1H-indole-2-carboxylic acid ethyl ester (1.62 g, 8.56 mmol) andpotassium carbonate (4.17 g, 30 mmol) in 100 mL of acetone was refluxedfor 1 day. After cooling to room temperature the reaction was quenchedby addition of water and then partitioned between water and methylenechloride. The organic phase was dried over MgSO₄, and concentrated toyield ethyl 1-(4-nitrobenzyl)-1H-indole-2-carboxylate as an off-whitesolid (2.0 g, 72%): ¹H NMR (DMSO-d₆) δ 1.27 (t, J=7.1 Hz, 3H), 4.26(q,J=7.1 Hz, 3H), 5.99(s, 2H), 7.18(t, J=7.0 Hz, 1H), 7.22 (d, J=9.0-Hz,2H), 7.34 (t, J=7.0 Hz, 1H), 7.43 (s, 1H), 7.58 (d, J=13.8 Hz, 1H), 7.76(d, J=13.8 Hz, 1H), 8.15 (d, J=9.0 Hz, 2H); MS (ESI) m/z 325 (MH⁺).

Step 2: A large excess of Raney® nickel was added in portions to astirred solution of ethyl 1-(4-nitrobenzyl)-1H-indole-2-carboxylate(0.65 g, 2 mmol), hydrazine (0.5 mL, 16 mmol) in 25 mL of ethanol. Afterstirring at room temperature for 2 hours the catalyst was then removedby filtering through a short pad of Celite®521. The filtrate wasconcentrated to give ethyl 1-(4-aminobenzyl)-1H-indole-2-carboxylate (MS(ESI) m/z 294.2 (MH⁺) as a gummy solid. The gummy solid was thendissolved in 15 mL of methylene chloride. To the solution was addeddiisopropylethylamine (0.5 mL, 2.9 mmol) and biphenyl 4-sulfonylchloride (0.5 g, 1.0 mmol). After being stirred at room temperatureovernight the reaction was concentracted and the residue wasre-dissolved in 15 mL of 2:1:1 THF/MeOH/water. Lithium hydroxidemonohydrate (0.2 g, 4.8 mmol) was added and the mixture was stirred atroom temperature overnight. Most of the organic solvents was removed andthe reaction mixture was made acidic (pH 6) with glacial acetic acid,and the solid was collected and purified by semi-preparative HPLC. Theproduct was collected based on UV absorption and concentrated to givethe title compound as a brown solid (0.11 g, 22%): ¹H NMR (DMSO-d₆) δ5.75 (s, 2H), 6.93 (d, J=8.9 Hz, 2H), 7.01 (d, J=8.9 Hz, 2H), 7.09 (t,J=8.0 Hz, 1H), 7.43 (t, J=8.0 Hz, 1H), 7.49 (s, 1H), 7.40-7.55 (m, 4H);7.65-7.80 (m, 3H), 7.75-7.82 (m, 4H), 10.30 (s, 1H), 12.93 (s, 1H); HRMScalcd for C₂₈H₂₃N₂O₄S: 483.1367; found (ESI⁺): 483.1374; Anal. calcd forC₂₈H₂₂N₂O₄S 0.2H₂O: C, 69.18; H, 4.64; N, 5.76. Found: C, 69.21; H,4.60; N, 5.78.

Example 4 Synthesis of1-(4-{[(4-tert-butylphenyl)sulfonyl]amino}benzyl)-1H-indole-5-carboxylicAcid

Step 1: 1-(4-Nitro-benzyl)-1H-indole-5-carboxylic acid methyl ester wasprepared from 1H-indole-5-carboxylic acid methyl ester and 4-nitrobenzylbromide followed the procedure of Example 3 Step 1 as a yellow poder: ¹HNMR (DMSO-d₆) δ 3.84 (s, 3H), 5.67 (s, 2H), 6.71 (dd, J=3.2, 0.8 Hz,1H), 7.40 (d, J=8.8 Hz, 2H), 7.54 (d, J=8.7 Hz, 1H), 7.67 (d, J=0.8 Hz,1H), 7.73 (dd, J=8.8, 1.7 Hz, 1H), 8.19 (d, J=8.8 Hz, 2H), 8.29 (d,J=1.0 Hz, 1H); MS (ESI) m/z 311 (MH⁺), 309 ([M−H]⁻); HRMS calcd forC₁₇H₁₅N₂O₄: 311.1030; found (ESI⁺): 311.1021.

Step 2: The title compound was prepared from1-(4-nitro-benzyl)-1H-indole-5-carboxylic acid methyl ester and4-tert-butylbenzene sulfonyl chloride followed the procedure of Example3 Step 2 as a light brown solid: ¹H NMR (DMSO-d₆) δ 1.24 (s, 9H), 5.33(s, 2H), 6.59 (d, J=3.2 Hz, 1H), 7.02 (d, J=8.7 Hz, 2H), 7.07 (d, J=8.7Hz, 2H), 7.45 (d, J=8.7 Hz, 1H), 7.51-7.54 (m, 3H), 7.65 (d, J=8.5 Hz,2H), 7.69 (d, J=1.7 Hz, 1H), 8.21 (d, J=1.3 Hz, 1H), 10.23 (br s, 1H),12.39 (br s, 1H); MS (ESI) m/z 463 (MH⁺), 461 ([M−H]⁻); HRMS calcd forC₂₆H₂₆N₂O₄S: 463.1690; found (ESI⁺): 463.1681.

Example 5 Synthesis of1-{4-[(1,1′-biphenyl-4-ylsulfonyl)amino]benzyl}-1H-indole-5-carboxylicAcid

The title compound was prepared from1-(4-nitro-benzyl)-1H-indole-5-carboxylic acid methyl ester andbisphenyl-4-sulfonyl chloride followed the procedure of Example 3 Step 2as a white solid: ¹H NMR (DMSO-d₆) δ 5.34 (s, 3H), 6.59 (d, J=3.1 Hz,1H), 7.06 (d, J=11.2 Hz, 2H), 7.10 (d, J=8.7 Hz, 2H), 7.40-7.48 (m, 4H),7.52 (d, J=3.1 Hz, 1H), 7.67-7.70 (m, 3H), 7.75-7.85 (m, 4H), 8.21 (d,J=1.4 Hz, 1H), 10.34 (br s, 1H), 12.37 (br s, 1H); MS (ESI) m/z 483(MH⁺), 481 ([M−H]⁻); HRMS calcd for C₂₈H₂₃N₂O₄S: 483.1376; found (ESI⁺):483.1368.

Example 6 Synthesis of1-(4-{[(4-tert-butylphenyl)sulfonyl]amino}benzyl)-1H-indole-3-carboxylicAcid

Step 1: 1-(4-Nitro-benzyl)-1H-indole-3-carboxylic acid methyl ester(113220-11) was prepared from 1H-indole-3-carboxylic acid methyl esterand 4-nitrobenzyl bromide followed the procedure of Example 3 Step 1 asa yellow powder: ¹H NMR (DMSO-d₆) δ 3.83 (s, 3H), 5.70 (s, 2H), 7.23(ddd, J=9.9, 4.6, 0.8 Hz, 2H), 7.47 (d, J=8.8 Hz, 2H), 7.50 (ddd, J=9.9,4.6, 0.8 Hz, 1H), 8.05 (ddd, J=9.9, 4.6, 0.8 Hz, 1H), 8.19 (d, J=8.8 Hz,2H), 8.40 (s, 1H); MS (ESI) m/z 311 (MH⁺), 309 ([M−H]⁻).

Step 2: The title compound was prepared from1-(4-nitro-benzyl)-1H-indole-3-carboxylic acid methyl ester and4-tert-butylbenzenesulfonyl chloride followed the procedure of Example 3Step 2 as a light brown solid: ¹H NMR (DMSO-d₆) δ 1.24 (s, 9H), 5.36 (s,2H), 7.04 (d, J=8.5 Hz, 2H), 7.13-7.18 (m, 4H), 7.45 (dd, J=6.1, 2.6 Hz,1H), 7.52 (d, J=8.8 Hz, 2H), 7.66 (d, J=8.8 Hz, 2H), 8.00 (dd, J=6.1,2.6 Hz, 1H), 8.13 (s, 1H), 10.23 (br s, 1H), 11.98 (br s, 1H); MS (ESI)m/z 463 (MH⁺), 461 ([M−H]⁻); HRMS calcd for C₂₆H₂₇N₂O₄S: 463.1690; found(ESI⁺): 463.1681; Anal. calcd for C₂₆H₂₆N₂O₄S-0.06 TFA: C, 67.51; H,5.67; N, 6.06. Found: C, 67.40; H, 5.65; N, 6.04.

Example 7 Synthesis of1-{4-[(1,1′-biphenyl-4-ylsulfonyl)amino]benzyl}-1H-indole-3-carboxylicAcid

The title compound was prepared from1-(4-nitro-benzyl)-1H-indole-3-carboxylic acid methyl ester andbisphenyl-4-sulfonyl chloride followed the procedure of Example 3 Step 2as an off-white solid: ¹H NMR (DMSO-d₆) δ 5.37 (s, 2H), 7.07 (d, J=8.7Hz, 2H), 7.13-7.18 (m, 4H), 7.40-7.51 (m, 4H), 7.67 (d, J=7.1 Hz, 2H),7.75-7.85 (m, 4H), 8.00 (dd, J=5.8, 0.7 Hz, 1H), 8.14 (s, 1H), 10.36 (brs, 1H), 12.01 (br s, 1H); MS (ESI) m/z 483 (MH⁺), 481 ([M−H]⁻); HRMScalcd for C₂₈H₂₃N₂O₄S, 483.1376; found (ESI⁺): 483.1367; Anal. calcd forC₂₈H₂₂N₂O₄S_(0.1) TFA: C, 68.57; H, 4.51; N, 5.67. Found: C, 68.67; H,4.50; N, 5.70.

Example 8 Synthesis of1-(4-{[(4-tert-butylphenyl)sulfonyl]amino}benzyl)-1H-indole-4-carboxylicAcid

Step 1: 1-(4-Nitro-benzyl)-1H-indole-4-carboxylic acid methyl ester wasprepared from 1H-indole-4-carboxylic acid methyl ester and 4-nitrobenzylbromide followed the procedure of Example 3 Step 1 as a yellow powder:¹H NMR (DMSO-d₆) δ 3.90 (s, 3H), 5.69 (s, 2H), 7.05(dd, J=3.1, 0.9 Hz,1H), 7.22 (t, J=7.6 Hz, 1H), 7.36 (d, J=8.9 Hz, 2H), 7.75 (td, J=7.6,3.1 Hz, 3H), 8.18 (d, J=8.9 Hz, 2H); MS (ESI) m/z 311 (MH⁺), 309([M−H]⁻); Anal. calcd for C₁₇H₁₄N₂O₄: C, 65.80; H, 4.55; N, 9.03. Found:C, 65.81; H, 4.50; N, 8.98.

Step 2: The title compound was prepared from1-(4-nitro-benzyl)-1H-indole-4-carboxylic acid methyl ester and4-tert-butylbenzenesulfonyl chloride followed the procedure of Example 3Step 2 as a pale pink solid: ¹H NMR (DMSO-d₆) δ 1.24 (s, 9H), 5.35 (s,2H), 6.96 (d, J=3.1 Hz, 1H), 7.03 (d, J=8.7 Hz, 2H), 7.07 (d, J=8.0 Hz,2H), 7.15 (t, J=7.8 Hz, 1H), 7.53 (d, J=8.8 Hz, 2H), 7.57 (d, J=3.2 Hz,1H), 7.62-7.68 (m, 3H), 7.70 (dd, J=7.5, 0.9 Hz, 1H), 10.26 (br s, 1H),12.59 (br s, 1H); MS (ESI) m/z 463 (MH⁺), 461 ([M−H]—); HRMS calcd forC₂₆H₂₇N₂O₄S: 463.1690; found (ESI⁺): 463.1674.

Example 9 Synthesis of1-{4-[(1,1′-biphenyl-4-ylsulfonyl)amino]benzyl}-1H-indole-4-carboxylicAcid

The title compound was prepared from1-(4-nitro-benzyl)-1H-indole-4-carboxylic acid methyl ester andbisphenyl-4-sulfonyl chloride followed the procedure of Example 3 Step 2as an off-white solid: ¹H NMR (DMSO-d₆) δ 5.36 (s, 2H), 6.96 (d, J=3.1Hz, 1H), 7.05 (d, J=8.0 Hz, 2H), 7.09 (d, J=9.0 Hz, 2H), 7.15 (t, J=7.8Hz, 1H), 7.40-7.47 (m, 1H), 7.49-7.51 (m, 2H), 7.57 (d, J=3.1 Hz, 1H),7.64-7.71 (m, 4H), 7.75-7.85 (m, 4H), 10.33 (br s, 1H), 12.58 (br s,1H); MS (ESI) nz/z 483 (MH⁺); 481 ([M−H]⁻); HRMS calcd for C₂₈H₂₃N₂O₄S:483.1376; found (ESI⁺): 483.1367.

Example 10 Synthesis of1-{4-[(1,1′-biphenyl-4-ylsulfonyl)amino]benzyl}-1H-indole-6-carboxylicAcid

Step 1: 1-(4-Nitrobenzyl)-1H-indole-6-carboxylic acid methyl ester wasprepared from 1H-indole-6-carboxylic acid methyl ester and 4-nitrobenzylbromide followed the procedure of Example 3 Step 1 as a yellow powder:¹H NMR (DMSO-d₆) δ 3.85 (s, 3H), 5.75 (s, 2H), 6.66(dd, J=3.1, 0.6 Hz,1H), 7.34 (d, J=8.7 Hz, 2H), 7.60 (t, J=2.8 Hz, 1H), 7.79 (d, J=3.1 Hz,1H), 8.07 (s, 1H), 8.15 (s, 1H), 8.19 (d, J=8.8 Hz, 1H), 8.28 (d, J=8.7Hz, 2H); MS (ESI) m/z 311 (MH⁺); 309 ([M−H]⁻).

Step 2: The title compound was prepared from1-(4-nitro-benzyl)-1H-indole-6-carboxylic acid methyl ester andbisphenyl-4-sulfonyl chloride followed the procedure of Example 3 Step 2as a light yellow solid: ¹H NMR (DMSO-d₆) δ 5.40 (s, 2H), 6.55(d, J=3.9Hz, 1H), 7.06 (s, 4H), 7.40-7.43 (m, 1H), 7.47-7.51 (m, 2H), 7.60 (d,J=1.2 Hz, 1H), 7.63 (d, J=3.1 Hz, 1H), 7.67 (dt, J=7.0, 1.2 Hz, 2H),7.75-7.85 (m, 4H), 7.99 (d, J=0.9 Hz, 1H), 10.32 (br s, 1H), 12.55 (brs, 1H); MS (ESI) m/z 483 (MH⁺); 481 ([M−H]⁻); HRMS calcd forC₂₈H₂₃N₂O₄S: 483.1376; found (ESI⁺): 483.1368; Anal. calcd forC₂₈H₂₂N₂O₄S_(0.02) TFA C, 69.21; H, 4.60; N, 5.76. Found: C, 69.01H,4.48 N, 5.78.

Example 11 Synthesis of3-phenyl-1-{4-[(phenylsulfonyl)amino]benzyl}-1H-indole-2-carboxylic Acid

Step 1: A solution of N-bromosuccinimide (4.94 g, 27.7 mmol) in DMF (20mL) was added dropwise to a stirred solution of ethyl1H-indole-2-carboxylate (5.0 g, 26.4 mmol) in DMF (15 mL) at 0˜5° C.After the addition was completed, the reaction was warmed to roomtemperature and stirring was continued for an additional 1.5 h. Thereaction mixture was poured over ice water and the resulting precipitatewas collected and dried to give 3-bromo-1H-indolecarboxylic acid ethylester (6.68 g, 81%) as a white solid: ¹H NMR (DMSO-d₆) δ 1.37 (t, J=7.2Hz, 3H), 4.38 (q, J=7.2 Hz, 2H), 7.18-7.22 (m, 1H), 7.34-7.38 (m, 1H),7.50 (dd, J=8.3, 0.8 Hz, 1H), 7.55 (dd, J=8.3, 0.8 Hz, 1H), 12.22 (br s,1H); MS (ESI) m/z 268 (MH⁺); 266 ([M−H]⁻).

Step 2: A mixture of 3-bromo-1H-indolecarboxylic acid ethyl ester (0.50g, 1.87 mmol), phenylboronic acid (0.27 g, 2.25 mmol), 2 M aqueoussodium carbonate (8 mL), tetrakis(triphenylphosphine)palladium (0) (0.20g, 0.17 mmol) in ethanol (16 mL) and toluene (30 ml) was heated at 65°C. for 16 h and then cooled. The reaction mixture was diluted with 1 Nhydrochloric acid and then extracted with ethyl acetate. The organicextracts were washed with water, dried over magnesium sulfate andconcentrated. Flash silica gel chromatography using ethyl acetate/hexaneas elutant gave 0.34 g (68%) of 3-phenyl-1H-indolecarboxylic acid ethylester as a pink solid: ¹H NMR (DMSO-d₄) δ 1.18 (t, J=7.2 Hz, 3H), 4.22(q, J=7.2 Hz, 2H), 7.07-7.11 (m, 1H), 7.29-7.33 (m, 1H) 7.45 (dd, J=8.3,0.8 Hz, 1H), 7.48-7.50 (m, 5H), 7.50 (dd, J=8.3, 0.8 Hz, 1H), 11.90 (brs, 1H); MS (ESI) m/z 266 (MH⁺); 264 ([M−H]⁻).

Step 3: 1-(4-Nitro-benzyl)-3-phenyl-1H-indolecarboxylic acid ethyl esterwas prepared from of 3-phenyl-1H-indolecarboxylic acid ethyl ester and4-nitrobenzyl bromide followed the procedure of Example 3 Step 1 as ayellow powder: ¹H NMR (DMSO-d₆) δ ¹H NMR (DMSO-d₆) δ 0.91 (t, J=7.2 Hz,3H), 4.05 (q, J=7.2 Hz, 2H), 5.97 (s, 2H), 7.17-7.23 (m, 1H), 7.32 (d,J=8.8 Hz, 2H), 7.36-7.42 (m, 1H), 7.38-7.42 (m, 1H), 7.45-7.48 (m, 4H),7.52 (d, J=8.2 Hz, 1H), 7.62 (d, J=8.2 Hz, 1H), 8.19 (d, J=8.8 Hz, 2H);MS (ESI) m/z 401 (MH⁺); 399 ([M−H]⁻); HRMS calcd for C₂₄H₂₁N₂O₄:401.1490; found (ESI⁺): 401.1494; Anal. calcd for C₂₄H₂₀N₂O₄.0.05 TFA C,71.99; H, 5.03; N, 7.00 Found: C, 71.27; H, 4.98; N, 6.90.

Step 4: 1-(4-Amino-benzyl)-3-phenyl-1H-indole indolecarboxylic acidethyl ester was prepared from1-(4-nitro-benzyl)-3-phenyl-1H-indolecarboxylic acid ethyl esterfollowed the procedure of Example 1 Step 1 as a yellow syrup: ¹H NMR(DMSO-d₆) δ 0.98 (t, J=7.2 Hz, 3H), 4.11 (q, J=7.2 Hz, 2H), 4.99 (br s,2H), 5.59 (s, 2H), 6.44 (d, J=8.41 Hz, 2H), 6.85 (d, J=8.4 Hz, 2H), 7.13(td, J=7.1, 0.8 Hz, 1H), 7.34 (td, J=7.1, 0.8 Hz, 1H), 7.38-7.41 (m,3H), 7.44-7.49 (m, 3H), 7.69 (d, J=8.4 Hz, 1H); MS (ESI) Hz 371 (MH⁺);369 ([M−H]⁻); Anal. calcd for C₂₄H₂₂N₂O₂.0.04 TFA C, 71.12; H, 5.92; N,7.47. Found: C, 71.03; H, 5.87; N, 7.36.

Step 5: The title compound was prepared from1-(4-amino-benzyl)-3-phenyl-1H-indole indolecarboxylic acid ethyl esterand benzenesulfonyl chloride followed the procedure of Example 3 Step 2as an off-white solid: ¹H NMR (DMSO-d₆) δ 5.72 (s, 2H), 6.95-7.00 (m,4H), 7.09-7.13 (m, 1H), 7.27-7.31 (m, 1H), 7.33-7.37 (m, 1H), 7.42-7.45(m, 5H), 7.49-7.55 (m, 3H), 7.57-7.61 (m, 1H), 7.72 (dd, J=7.1, 1.4 Hz,2H), 10.26 (br s, 1H), 12.83 (br s, 1H); MS (ESI) m/z 481 ([M−H]⁻); HRMScalcd for C₂₈H₂₁N₂O₄S: 481.1224; found (ESI⁻): 481.1228.

Example 12 Synthesis of1-{4-[(methylsulfonyl)amino]benzyl}-3-phenyl-1H-indole-2-carboxylic Acid

The title compound was prepared from1-(4-nitro-benzyl)-3-phenyl-1H-indole indolecarboxylic acid ethyl esterand methanesulfonyl chloride followed the procedure of Example 3 Step 2as an off-white solid: ¹H NMR (DMSO-d₆) δ 2.94 (s, 3H), 5.80 (s, 2H),7.05-7.15 (m, 5H), 7.31-7.38 (m, 2H), 7.43-7.47 (m, 5H), 7.62 (d, J=8.4Hz, 1H), 9.68 (br s, 1H), 12.87 (br s, 1H); MS (ESI) nz 419 ([M−H]⁻);HRMS calcd for C₂₃H₁₉N₂O₄S: 419.1068; found (ESI⁻): 419.1074.

Example 13 Synthesis of1-{4-[(phenylsulfonyl)amino]benzyl}-3-thien-2-yl-1H-indole-2-carboxylicAcid

Step 1: 3-Thiophene-2-yl-1H-indole-2-carboxylic acid ethyl ester wasprepared from 3-bromo-1H-indolecarboxylic acid ethyl ester and2-thiopheneboronic acid followed the procedure of Example 11 Step 2 as alight yellow powder: ¹H NMR (DMSO-d₆) δ 1.27 (t, J=7.2 Hz, 3H), 4.30 (q,J=7.2 Hz, 2H), 7.12-7.16 (m, 1H), 7.17-7.20 (m, 1H), 7.31-7.34 (m, 1H),7.36(dd, J=5.1, 1.5 Hz, 1H), 7.51 (d, J=8.3 Hz, 1H), 7.63 (d, J=6.3 Hz,1H), 7.71 (d, J=8.2 Hz, 1H), 12.02 (br s, 1H); MS (ESI) m/z 272 (MH⁺);270 ([M−H]⁻).

Step 2: 1-(4-Nitro-benzyl)-3-thiophene-2-yl-1H-indole-2-carboxylic acidethyl ester was prepared from 3-thiophene-2-yl-1H-indole-2-carboxylicacid ethyl ester and 4-nitrobenzyl bromide followed the procedure ofExample 3 Step 1 as a yellow powder: ¹H NMR (DMSO-d₆) δ 1.02 (t, J=7.2Hz, 3H), 4.15 (q, J=7.2 Hz, 2H), 7.20 (dd, J=6.9, 3.4 Hz, 1H), 7.24 (dd,J=6.2, 0.8 Hz, 1H), 7.25-7.27 (m, 1H), 7.31 (d, J=8.8 Hz, 2H), 7.37-7.43(m, 1H), 7.62 (d, J=8.4 Hz, 1H), 7.68 (dd, J=5.2, 1.2 Hz, 1H), 7.71 (d,J=4.1 Hz, 1H), 8.19 (d, J=8.8 Hz, 2H); MS (ESI) m/z 407 (MH⁺); 405([M−H]⁻); Anal. calcd for C₂₂H₁₈N₂O₄S: C, 65.01; H, 4.46; N, 6.89 Found:C, 65.04; H, 6.62; N, 6.82.

Step 3: 1-(4-Amino-benzyl)-3-thiophene-2-yl-1H-indole-2-carboxylic acidethyl ester was prepared from1-(4-nitro-benzyl)-3-thiophene-2-yl-1H-indole-2-carboxylic acid ethylester followed the procedure of Example 1 Step 1: ¹H NMR (DMSO-d₆) δ1.10 (t, J=7.2 Hz, 3H), 4.20 (q, J=7.2 Hz, 2H), 5.01 (br s, 2H), 5.55(s, 2H), 6.45 (d, J=8.5 Hz, 2H), 6.84 (d, J=8.5 Hz, 2H), 7.16-7.20 (m,3H), 7.35 (td, J=7.3, 1.0 Hz, 1H), 7.62 (dd, J=5.2, 1.2 Hz, 1H), 7.65(d, J=4.1 Hz, 1H), 7.71 (d, J=8.8 Hz, 1H); MS (ESI) m/z 377 (MH⁺); 375([M−H]⁻); Anal. calcd for C₂₂H₂₀N₂O₂S-0.05 TFA C, 69.46; H, 5.29; N,7.33. Found: C, 69.66; H, 5.04; N, 7.72.

Step 4: The title compound was prepared from1-(4-amino-benzyl)-3-thiophene-2-yl-1H-indole-2-carboxylic acid ethylester and benzenesulfonyl chloride followed the procedure of Example 3Step 2 as a light brown solid: ¹H NMR (DMSO-d₆) δ 5.67 (s, 2H),6.95-7.05 (m, 4H), 7.14-7.23 (m, 3H), 7.30-7.35 (m, 1H), 7.49-7.75 (m,8H), 10.26 (br s, 1H), 13.24 (br s, 1H); MS (ESI) m/z 489 (MH⁺); HRMScalcd for C₂₆H₂₁N₂O₄S₂: 489.0940; found (ESI⁺): 489.0935.

Example 14 Synthesis of1-{4-[(methylsulfonyl)amino]benzyl}-3-thien-2-yl-1H-indole-2-carboxylicAcid

The title compound was prepared from1-(4-nitro-benzyl)-3-thiophene-2-yl-1H-indole-2-carboxylic acid ethylester and methanesulfonyl chloride followed the procedure of Example 3Step 2 as a light yellow solid: ¹H NMR (DMSO-d₆) δ 2.94 (s, 3H), 5.76(s, 2H), 7.08-7.24 (m, 7H), 7.34 (td, J=8.4, 1.2 Hz, 1H), 7.60-7.70 (m,3H), 9.69 (br s, 1H), 13.25 (br s, 1H); MS (ESI) m/z 427 (MH⁺); MS (ESI)m/z 425 ([M−H]⁻); HRMS calcd for C₂₁H₁₉N₂O₄S₂: 427.0784; found (ESI⁺):427.0779.

Example 15 Synthesis of1-[4-(2,5-dimethyl-1H-pyrrol-1-yl)benzyl]-3-phenyl-1H-indole-2-carboxylicAcid

A mixture of 1-(4-amino-benzyl)-3-phenyl-1H-indolecarboxylic acid ethylester (0.20 g, 0.54 mmol), acetonylacetone (0.16 mL, 1.35 mmol), andtoluene (15 mL) was heated at reflux under nitrogen using a Dean-Starktrap for 6 h. The reaction was cooled and concentrated. The residue wasre-dissolved in 10 mL of 2:1:1 THF/MeOH/water. Lithium hydroxidemonohydrate (0.09 g, 2.1 mmol) was added and the mixture was stirred atroom temperature overnight. Most of the organic solvents was removed andthe reaction mixture was made acidic (pH 6) with glacial acetic acid,and the solid was collected and purified by HPLC to afford the titlecompound (0.027 g, 6%) as a brown solid: ¹H NMR (DMSO-d₆) δ 1.91 (s,6H), 5.75 (s, 2H), 7.01-7.30 (m, 6H), 7.32-7.38 (m, 2H), 7.40-7.50 (m,6H), 7.68 (d, J=8.54 Hz, 1H), 12.80 (br s, 1H); MS (ESI) m/z 421 (MH⁺);MS (ESI) m/z 419 ([M−H]⁻); HRMS calcd for C₂₈H₂₆N₂O₂: 421.1905; found(ESI⁺): 421.1915.

Example 16 Synthesis of1-(4-anilinobenzyl)-3-phenyl-1H-indole-2-carboxylic Acid

Phenylboronic acid (0.16 g, 0.81 mmol), Cu(OAc)₂ (0.049 g, 0.27 mmol),and myristic acid (0.037 g, 0.16 mmol) were combined in a 100-mLround-bottom flask with a large stir bar. A rubber septum was attached,and dry toluene (2 mL), 2,6-lutidine (0.066 mL, 0.57 mmol), and1-(4-amino-benzyl)-3-phenyl-1H-indole indolecarboxylic acid ethyl ester(0.20 g, 0.54 mmol) were successively added. The resulting mixture wasstirred at a high rate for 24 h, diluted with ethyl acetate (10 mL),filtered through a plug of silica gel, and then purified by columnchromatography (5 to 25% EtOAc/hexanes) to give a gummy solid. To astirred solution of the gummy solid in 15 mL of 2:1:1 THF/MeOH/water wasadded lithium hydroxide monohydrate (0.1 g, 2.4 mmol). The reaction wasstirred at 40° C. for 5 h. Most of the organic solvents was removed andthe reaction mixture was made acidic (pH 6) with glacial acetic acid,and the solid was collected and purified by HPLC as a white powder(0.023 g, 10%): ¹H NMR (DMSO-d₆) δ 5.64 (s, 2H), 6.77 (t, J=7.4 Hz, 1H),6.96 (d, J=8.5 Hz, 2H), 7.00 (dd, J=7.6, 0.9 Hz, 2H), 7.04(t, J=7.1 Hz,1H), 7.15-7.19(m, 4H), 7.25 (t, J=7.11 Hz, 1H), 7.39 (t, J=7.4 Hz, 2H),7.46-7.53 (m, 1H), 7.53 (t, J=7.4 Hz, 2H), 7.54-7.56 (m, 2H), 8.08 (brs, 1H), 13.01 (br s, 1H); MS (ESI) m/z 419 (MH⁺); MS (ESI) m/z 417([M−H]⁻); HRMS calcd for C₂₈H₂₃N₂O₂: 419.1757; found (ESI⁺): 419.1755.

Example 17 Synthesis of1-(4-anilinobenzyl)-3-thien-2-yl-1H-indole-2-carboxylic Acid

The title compound was prepared from1-(4-amino-benzyl)-3-thiophene-2-yl-1H-indole-2-carboxylic acid ethylester and phenylboronic acid followed the procedure of Example 16 as alight green solid: ¹H NMR (DMSO-d₆) δ 5.70 (s, 2H), 6.78 (td, J=7.4, 1.0Hz, 1H), 6.98 (d, J=8.7 Hz, 2H), 7.01 (dd, J=8.5, 1.0 Hz, 2H), 7.04 (d,J=8.7 Hz, 2H), 7.16 (t, J=3.5 Hz, 1H), 7.17 (dd, J=5.2, 1.6 Hz, 2H),7.21 (dd, J=6.9, 2.1 Hz, 2H), 7.33-7.37 (m, 1H), 7.61 (dd, J=5.1, 1.3Hz, 1H), 7.66 (t, J=8.7 Hz, 2H), 8.12 (br s, 1H), 13.34 (br s, 1H); MS(ESI) m/z 425 (MH⁺); MS (ESI) m/z 423 ([M−H]⁻); HRMS calcd forC₂₆H₂₁N₂O₂S: 425.1321; found (ESI⁺): 425.1319.

Example 18 Primary Screen for the PAI-1 Inhibition

Test compounds are dissolved in DMSO at a final concentration of 10 mM,then diluted 100×in physiologic buffer. The inhibitory assay isinitiated by the addition of the test compound (1-100 μM finalconcentration, maximun DMSO concentration of 0.2%) in a pH 6.6 buffercontaining 140 nM recombinant human plasminogen activator inhibitor-1(PAI-1; Molecular Innovations, Royal Oak, Mich.). Following a 1 hourincubation at room temperature, 70 nM of recombinant human tissueplasminogen activator (tPA) is added, and the combination of the testcompound, PAI-1 and tPA is incubated for an additional 30 minutes.Following the second incubation, Spectrozyme-tPA (American Diagnostica,Greenwich, Conn.), a chromogenic substrate for tPA, is added andabsorbance read at 405 nm at 0 and 60 minutes. Relative PAI-1 inhibitionis equal to the residual tPA activity in the presence of the testcompound and PAI-1. Control treatments include the complete inhibitionof tPA by PAI-1 at the molar ratio employed (2:1), and the absence ofany effect of the test compound on tPA alone.

Example 19 Assay for Determining IC₅₀ of Inhibition of PAI-1

This assay is based upon the non-SDS dissociable interaction between tPAand active PAI-1. Assay plates are initially coated with human tPA (10μg/ml). Test compounds of the present invention are dissolved in DMSO at10 mM, then diluted with physiologic buffer (pH 7.5) to a finalconcentration of 1-50 μM. Test compounds are incubated with human PAI-1(50 ng/ml) for 15 minutes at room temperature. The tPA-coated plate iswashed with a solution of 0.05% Tween 20 and 0.1% BSA, then the plate isblocked with a solution of 3% BSA. An aliquot of the substitutedindole/PAI-1 solution is then added to the tPA-coated plate, incubatedat room temperature for 1 hour, and washed. Active PAI-1 bound to theplate is assessed by adding an aliquot of a 1:1000 dilution of the 33B8monoclonal antibody against human PAI-1, and incubating the plate atroom temperature for 1 hour (Molecular Innovations, Royal Oak, Mich.)The plate is again washed, and a solution of goat anti-mouseIgG-alkaline phosphatase conjugate is added at a 1:50,000 dilution ingoat serum. The plate is incubated 30 minutes at room temperature,washed, and a solution of alkaline phosphatase substrate is added. Theplate is incubated 45 minutes at room temperature, and color developmentis determined at OD_(405 nm). The quantitation of active PAI-1 bound totPA at varying concentrations of the test compound is used to determinethe IC₅₀. Results are analyzed using a logarithmic best-fit equation.The assay sensitivity is 5 ng/ml of human PAI-1 as determined from astandard curve ranging from 0-100 ng/ml.

The compounds of the present invention inhibited Plasminogen ActivatorInhibitor-1 as summarized in Table 1. TABLE 1 Compound IC₅₀ % Inhibition@ 10 % Inhibition @ 25 Number μM μM μM 1 39 40 2 15 35 3 66 4 27 5 22 619 28 7 25 27 8 27 47 9 26 35 10 26 30 11 51 71 12 42 49 13 50 79 14 4553 15 2.51 53 69 16 26 32 17 25 35

Although the foregoing invention has been described in detail by way ofexample for purposes of clarity of understanding, it will be apparent tothe artisan that certain changes and modifications are comprehended bythe disclosure and can be practiced without undue experimentation withinthe scope of the appended claims, which are presented by way ofillustration not limitation.

All publications and patent documents cited above are herebyincorporated by reference in their entirety for all purposes to the sameextent as if each were so individually denoted.

1. A compound having the formula:

or a pharmaceutically acceptable salt or ester form thereof, wherein: Xis a bond or C₁-C₆ alkylene; R₁ and R₂ are, independently, hydrogen,C₁-C₆ alkyl, C₂-C₇ alkenyl, C₂-C₇ alkynyl, C₃-CS cycloalkyl, C₇-C₁₁bicycloalkyl, C₆-C₁₀ aryl, heterocycle, —C(═O)C₁-C₆ alkyl, —C(═O)aryl,—C(═O)heterocycle, —C(═O)N(R₆)C₁-C₆ alkyl, —C(═O)N(R₆)aryl,—C(═O)N(R₆)heterocycle, —SO₂—C₁-C₆ alkyl, —SO₂-aryl, or—SO₂-heterocycle; or R₁ and R₂ together form a heterocycle; R₃ and R₄are independently, hydrogen, halogen, C₁-C₆ alkyl, C₁-C₃ perfluoroalkyl,C₁-C₆ alkoxy, C₃-C₈ cycloalkyl, —C(═O)C₁-C₃ alkyl, —OH, —NH₂, or —NO₂;R₅ is hydrogen, C₁-C₆ alkyl, C₂-C₇ alkenyl, C₂-C₇ alkynyl, C₃-C₈cycloalkyl, C₇-C₁₁ bicycloalkyl, C₆-C₀ aryl, or heterocycle; A whenpresent is hydrogen, C₆-C₁₀ aryl, or heterocycle; and R₆ is hydrogen,C₁-C₆ alkyl, halogen, C₂-C₇ alkenyl, C₂-C₇ alkynyl, C₃-C₈ cycloalkyl,aralkyl, C₆-C₁₀ aryl, heterocycle, hydroxy, C₁-C₆ alkoxy, aryl-oxy, oxo(═O), —CN, —C(═O)H, —CO₂H, —OCO₂C₁-C₆ alkyl, —CO₂C₁-C₆ alkyl, —CO₂-aryl,—CO₂(C₁-C₆ alkyl)aryl, —OCO₂-aryl, —C(═O)NH₂, —C(═O)NHOH, amino, C₁-C₆alkylamino, dialkylamino of 1-6 carbons per alkyl moiety,—NHC(═O)NH—C₁-C₆ alkyl, —NHSO₂—C₁-C₆ alkyl, —NHSO₂-aryl, or—NHSO₂-heterocycle; and wherein each of said alkyl, alkoxy, alkenyl,alkynyl, cycloalkyl, aryl and heterocycle groups being optionallysubstituted by one, two, three or more substituents.
 2. A compound ofclaim 1, having the formula:

or a pharmaceutically acceptable salt of ester form thereof, wherein Yis NHSO₂—C₁-C₆ alkyl, NHSO₂-aryl, NHSO₂-heterocycle, pyrrole, NH-aryl,or NH-heterocycle.
 3. A compound of claim 1 wherein R₁ and R₂ are,independently, hydrogen, unsubstituted C₁-C₆ alkyl, C₂-C₇ alkenyl, C₂-C₇alkynyl, C₃-C₈ cycloalkyl, C₇-C₁₁ bicycloalkyl, C₆-C₁₀ aryl, heterocyle,—C(═O)C₁-C₆ alkyl, —C(═O)aryl, —C(═O)heterocycle, —C(═O)N(R₆)C₁-C₆alkyl, —C(═O)N(R₆)aryl, —C(═O)N(R₆)heterocycle, —SO₂—C₁-C₆ alkyl,—SO₂-aryl, or —SO₂-heterocycle.
 4. A compound of claim 1 wherein R₁ andR₂ are, independently, hydrogen, unsubstituted SO₂-alkyl, unsubstitutedSO₂-aryl, or SO₂-aryl optionally substituted with —OCF₃, aryl or alkyl.5. A compound of claim 1 wherein R₁ and R₂ together with the nitrogenform a heterocycle.
 6. A compound of claim 5 wherein R₁ and R₂ togetherwith the nitrogen form a pyrrole optionally substituted with one or twoC₁-C₆ alkyl groups.
 7. A compound of claim 1 wherein R₃ and R₄ are,independently, hydrogen, halogen, C₁-C₃ perfluoroalkyl, C₁-C₆ alkoxy,C₃-C₈ cycloalkyl, —C(═O)C₁-C₃ alkyl, —OH, —NH₂, —NO₂, unsubstitutedC₁-C₆ alkyl or C₁-C₆ alkyl substituted with halogen, —CN, or alkoxy. 8.A compound of claim 1 wherein R₅ is hydrogen.
 9. A compound of claim 1wherein A when present is hydrogen, phenyl, or thiophene.
 10. A compoundof claim 1 that is one of the following:1-[4-({[4-(trifluoromethoxy)phenyl]sulfonyl}amino)phenyl]-1H-indole-2-carboxylicacid or a pharmaceutically acceptable salt or ester form thereof;1-[4-(1H-pyrrol-1-yl)phenyl]-1H-indole-2-carboxylic acid or apharmaceutically acceptable salt or ester form thereof;1-{4-[(1,1′-biphenyl-4-ylsulfonyl)amino]benzyl}-1H-indole-2-carboxylicacid or a pharmaceutically acceptable salt or ester form thereof;1-(4-{[(4-tert-butylphenyl)sulfonyl]amino}benzyl)-1H-indole-5-carboxylicacid or a pharmaceutically acceptable salt or ester form thereof or1-{4-[(1,1′-biphenyl-4-ylsulfonyl)amino]benzyl}-1H-indole-5-carboxylicacid or a pharmaceutically acceptable salt or ester form thereof.
 11. Acompound of claim 1 that is that is one of the following:1-(4-{[(4-tert-butylphenyl)sulfonyl]amino}benzyl)-1H-indole-3-carboxylicacid or a pharmaceutically acceptable salt or ester form thereof;1-{4-[(1,1′-biphenyl-4-ylsulfonyl]amino}benzyl)-1H-indole-3-carboxylicacid or a pharmaceutically acceptable salt or ester form thereof;1-(4-{[(4-tert-butylphenyl)sulfonyl]amino}benzyl)-1H-indole-4-carboxylicacid or a pharmaceutically acceptable salt or ester form thereof;1-{4-[(1,1′-biphenyl-4-ylsulfonyl]amino}benzyl)-1H-indole-4-carboxylicacid or a pharmaceutically acceptable salt or ester form thereof or1-{4-[(1,1′-biphenyl-4-ylsulfonyl)amino]benzyl}-1H-indole-6-carboxylicacid or a pharmaceutically acceptable salt or ester form thereof.
 12. Acompound of claim 1 that is one of the following:3-phenyl-1-{4-[(phenylsulfonyl)amino]benzyl}-1H-indole-2-carboxylic acidor a pharmaceutically acceptable salt or ester form thereof;1-{4-[(methylsulfonyl)amino]benzyl}-3-phenyl-1H-indole-2-carboxylic acidor a pharmaceutically acceptable salt or ester form thereof;1-{4-[(phenylsulfonyl)amino]benzyl}-3-thien-2-yl-1H-indole-2-carboxylicacid or a pharmaceutically acceptable salt or ester form thereof;1-{4-[(methylsulfonyl)amino]benzyl}-3-thien-2-yl-1H-indole-2-carboxylicacid or a pharmaceutically acceptable salt or ester form thereof or1-[4-(2,5-dimethyl-1H-pyrrol-1-yl)benzyl]-3-phenyl-1H-indole-2-carboxylicacid or a pharmaceutically acceptable salt or ester form thereof.
 13. Acompound of claim 1 that is1-(4-anilinobenzyl)-3-phenyl-1H-indole-2-carboxylic acid or apharmaceutically acceptable salt or ester form thereof.
 14. A compoundof claim 1 that is1-(4-anilinobenzyl)-3-thien-2-yl-1H-indole-2-carboxylic acid or apharmaceutically acceptable salt or ester form thereof.
 15. A method ofinhibiting PAI-1 activity comprising administering to a subject in needthereof a therapeutically effective amount of a compound of claim
 1. 16.A method for treating a PAI-1 related disorder comprising administeringto a subject in need thereof a therapeutically effective amount of acompound of claim
 1. 17. The method of claim 16, wherein the PAI-1related disorder is impairment of the fibrinolytic system.
 18. Themethod of claim 16, wherein the PAI-1 related disorder is thrombosis,atrial fibrillation, pulmonary fibrosis, myocardial ischemia, stroke,thromboembolic complication of surgery, cardiovascular disease,atherosclerotic plaque formation, chronic obstructive pulmonary disease,renal fibrosis, polycystic ovary syndrome, diabetes, Alzheimer'sdisease, or cancer.
 19. The method of claim 19, wherein the thrombosisis selected from the group consisting of venous thrombosis, arterialthrombosis, cerebral thrombosis, and deep vein thrombosis.
 20. Themethod of claim 18, wherein the PAI-1 related disorder is cardiovasculardisease caused by noninsulin dependent diabetes mellitus in a subject.21. The method of claim 1, wherein the therapeutically effective amountis from 25 mg/kg/day to 200 mg/kg/day.
 22. A pharmaceutical compositioncomprising a compound of claim 1, or a pharmaceutically acceptable saltor ester form thereof, and a pharmaceutically acceptable excipient orcarrier.
 23. A method for treating thrombosis, atrial fibrillation,pulmonary fibrosis, thromboembolic complication of surgery, stroke,myocardial ischemia, atherosclerotic plaque formation, chronicobstructive pulmonary disease, or renal fibrosis comprisingadministering to a subject in need thereof a therapeutically effectiveamount of a compound of claim 1.